This document summarizes a seminar presentation on structural biology of nucleocytoplasmic transport. Some key points:
- Nucleocytoplasmic transport through the nuclear pore complex allows movement of RNA and proteins between the nucleus and cytoplasm.
- The nuclear pore complex regulates transport and consists of an elaborate multiprotein structure.
- Transport mechanisms involve nuclear localization signals, nuclear export sequences, transport receptors, and the RanGTPase cycle which establishes directionality.
- RNA transport mechanisms export messenger RNA, transfer RNA, ribosomal RNA, and reimport some nuclear RNAs through the nuclear pore complex.
- Future areas of study include better understanding transport receptor recognition and mRNA export pathways in
This presentation is about the transcription machinery that is required for the transcription in eukaryotes. The comparison between the transcription factors involved in prokaryotes and eukaryotes. The initiation of transcription and how it helps in producing a mRNA.
A detail ppt about Genome organization with focus on all levels of organization. Most recent research and findings about CT is also added in this ppt. Detail account of 30nm fiber and its ultra structure and types is also included.
This presentation is about the transcription machinery that is required for the transcription in eukaryotes. The comparison between the transcription factors involved in prokaryotes and eukaryotes. The initiation of transcription and how it helps in producing a mRNA.
A detail ppt about Genome organization with focus on all levels of organization. Most recent research and findings about CT is also added in this ppt. Detail account of 30nm fiber and its ultra structure and types is also included.
RNA Polymerase
Introduction
Purification
History
PRODUCTS OF RNAP
Messenger RNA
Non-coding RNA or "RNA genes
Transfer RNA
Ribosomal RNA
Micro RNA
Catalytic RNA (Ribozyme)
prokaryotic and eukaryotic
Transcription by RNA Polymerase
TYPES OF RNA POLYMERASE
Type I
Type II
Type III
Prokaryotic Transcription Unit
EXPRESSION OF A PROKARYOTIC GENE
Prokaryotic Polycistronic Message Codes for Several Different Proteins
Eukaryotic Transcription Unit
ENHANCERS AND SILENCERS
RESULT OF THE TRANSCRIPTION CYCLE
RNAP III TRANSCRIBES HUMAN MICRORNAS
RNAP I–specific subunits promotepolymerase clustering to enhance the rRNA genetranscription cycle
RNAP II–TFIIB STRUCTURE ANDMECHANISM OF TRANSCRIPTION INITIATION
FIVE CHECKPOINTS MAINTAINING THE FIDELITY OFTRANSCRIPTION BY RNAP IN STRUCTURAL ANDENERGETIC DETAILS
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Most bacteria are free-living organisms that grow by increasing
in mass and then divide by binary fission.
Growth and division are controlled by genes, the expression
of which must be regulated appropriately. Genes
whose activity is controlled in response to the needs of a
cell or organism are called regulated genes. All organisms
also have a large number of genes whose products
are essential to the normal functioning of a growing and
dividing cell, no matter what the conditions are. These
genes are always active in growing cells and are known as
constitutive genes or housekeeping genes; examples include
genes that code for the enzymes needed for protein
synthesis and glucose metabolism. Note that all genes are
regulated on some level. If normal cell function is impaired
for some reason, the expression of all genes, including
constitutive genes, is reduced by regulatory
mechanisms. Thus, the distinction between regulated
and constitutive genes is somewhat arbitrary.
An Overview...
Definition of Translation.
Def. of Eukaryotes.
Translation: An Overview.
Components of Translation.
Some Enzymes .
Ribosome Role.
Mechanism of Translation.
Initiation.
Scanning Model of Initiation.
Initiation Factors.
Animation.
Elongation.
Chain Elongation: Translocation.
Animation.
Termination.
Animation....
It's not perfect still... what are your views friends?
RNA transport
Multiple classes of RNA are exported from the nucleus
Transportation through nuclear pore complex.
Ribosomal subunits are assembled in the nucleolus and exported by exportin 1
tRNAs are exported by a dedicated exportin
Messenger RNAs are exported from the nucleus as RNA-protein complexes
Messenger RNAs are exported from the nucleus as RNA-protein complexes
hnRNPs move from sites of processing to NPCs
Precursors to microRNAs are exported from the nucleus and processed in the cytoplasm
RNA Polymerase
Introduction
Purification
History
PRODUCTS OF RNAP
Messenger RNA
Non-coding RNA or "RNA genes
Transfer RNA
Ribosomal RNA
Micro RNA
Catalytic RNA (Ribozyme)
prokaryotic and eukaryotic
Transcription by RNA Polymerase
TYPES OF RNA POLYMERASE
Type I
Type II
Type III
Prokaryotic Transcription Unit
EXPRESSION OF A PROKARYOTIC GENE
Prokaryotic Polycistronic Message Codes for Several Different Proteins
Eukaryotic Transcription Unit
ENHANCERS AND SILENCERS
RESULT OF THE TRANSCRIPTION CYCLE
RNAP III TRANSCRIBES HUMAN MICRORNAS
RNAP I–specific subunits promotepolymerase clustering to enhance the rRNA genetranscription cycle
RNAP II–TFIIB STRUCTURE ANDMECHANISM OF TRANSCRIPTION INITIATION
FIVE CHECKPOINTS MAINTAINING THE FIDELITY OFTRANSCRIPTION BY RNAP IN STRUCTURAL ANDENERGETIC DETAILS
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Most bacteria are free-living organisms that grow by increasing
in mass and then divide by binary fission.
Growth and division are controlled by genes, the expression
of which must be regulated appropriately. Genes
whose activity is controlled in response to the needs of a
cell or organism are called regulated genes. All organisms
also have a large number of genes whose products
are essential to the normal functioning of a growing and
dividing cell, no matter what the conditions are. These
genes are always active in growing cells and are known as
constitutive genes or housekeeping genes; examples include
genes that code for the enzymes needed for protein
synthesis and glucose metabolism. Note that all genes are
regulated on some level. If normal cell function is impaired
for some reason, the expression of all genes, including
constitutive genes, is reduced by regulatory
mechanisms. Thus, the distinction between regulated
and constitutive genes is somewhat arbitrary.
An Overview...
Definition of Translation.
Def. of Eukaryotes.
Translation: An Overview.
Components of Translation.
Some Enzymes .
Ribosome Role.
Mechanism of Translation.
Initiation.
Scanning Model of Initiation.
Initiation Factors.
Animation.
Elongation.
Chain Elongation: Translocation.
Animation.
Termination.
Animation....
It's not perfect still... what are your views friends?
RNA transport
Multiple classes of RNA are exported from the nucleus
Transportation through nuclear pore complex.
Ribosomal subunits are assembled in the nucleolus and exported by exportin 1
tRNAs are exported by a dedicated exportin
Messenger RNAs are exported from the nucleus as RNA-protein complexes
Messenger RNAs are exported from the nucleus as RNA-protein complexes
hnRNPs move from sites of processing to NPCs
Precursors to microRNAs are exported from the nucleus and processed in the cytoplasm
How are macromolecules transported across the nuclear envelope Desc.pdfkellenaowardstrigl34
How are macromolecules transported across the nuclear envelope? Describe the structure that
regulates nuclear membrane permeability.
Solution
The transportation of molecules in and out of the nucleus is controlled by the nuclear pore
complex(NPC). Small molecules move through the nuclear membrane with ease but for
macromolecules like proteins, RNA etc, they require an association with transportation factors
like importins, karyopherins and exportins.
Proteins that need to be carried into the nucleus have nuclear localization signals (NLS) which
bind to the importins. NLS is a sequence of Amino acids which act as a tag. They are mostly
hydrophilic but hydrophobic sequences have also been documented.
Nuclear Import:
Importins bind to the macromolecule to be transported in the cytoplasm itself. The complex then
interacts with the NPC and passes through the channel. Once inside the nucleus, it interacts with
Ran-GTP and the importin dissociates from the macromolecule it was carrying. Then the
importin-Ran-GTP complex is transported to the cytoplasm where it is separated from importin
by the Ran Binding Protein (RanBP). After being separated from importin, GTPase activating
protein(GAP) binds with Ran-GTP and induces hydrolysis of GTP to GDP. The Ran-GDP
produced binds with the nuclear transport factor(NUTF2) and then returns to the nucleoplasm.
Inside the nucleus, Ran-GDP interacts with guanine nucleotide exchange factor(GEF) which
replaces GDP with GTP and Ran-GTP is born again to repeat the cycle.
Nuclear Export:
In the nucleoplasm, exportin binds the macromolecule to be transported outside the nucleus and
Ran-GTP. The complex diffuses through the pore to the cytoplasm where it dissociates. Ran-
GTP binds to GAP and hydrolysis resulting in Ran-GDP. Ran-GDP is returned to the nucleus
where it exchanges its ligand for GTP.
Nuclear Pore:
The nuclear pore complex is made of proteins called nucleoporins. Around 50% of nucleoporins
contains solenoid protein domains (alpha or beta-propeller fold) and the rest might have both as
separate structural domains. Each NPS has about 456 individual protein molecules with 30
distinct nucleoproteins. These are highly flexible proteins that lack an ordered secondary
structure..
RNA splicing, in molecular biology, is a form of RNA processing in which a newly made precursor messenger RNA transcript is transformed into a mature messenger RNA. During splicing, introns are removed and exons are joined together.
For MBBS, BDS and General Biochemistry students, coding strand, sense strand, anti-sense strand, promoter, enhancers, silencers, TATA box, Goldberg Hogness box, alternative spilicing, post-transcriptional modification
Transcription in eukariotes by kk sahuKAUSHAL SAHU
INTRODUCTION
A STRUCTURAL GENE
EUKARYOTIC RNAPs
MACHANISM OF TRANSCRIPTION IN EUKARYOTES:
- INITIATION
-ELONGATION
-TERMINATION
RNA SPLISING
DIFFERENT BETWEEN PROKARYOTIC & EUKARYOTIC TRANSCRIPTION
BIBLIOGRAPHY
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Normal Labour/ Stages of Labour/ Mechanism of LabourWasim Ak
Normal labor is also termed spontaneous labor, defined as the natural physiological process through which the fetus, placenta, and membranes are expelled from the uterus through the birth canal at term (37 to 42 weeks
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
for beginners, providing thorough training in areas such as SEO, digital communication marketing, and PPC training in Noida. After finishing the program, students receive the certifications recognised by top different universitie, setting a strong foundation for a successful career in digital marketing.
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Introduction to AI for Nonprofits with Tapp NetworkTechSoup
Dive into the world of AI! Experts Jon Hill and Tareq Monaur will guide you through AI's role in enhancing nonprofit websites and basic marketing strategies, making it easy to understand and apply.
3. Flow of seminar
Introduction
Nuclear pore complex
Structure of nuclear pore complex
Mechanism of import and export of protiens
Rangtpase cycle
Rna transport
Research evidences
conclusion
4. What is NucleoCytoplasmic Transport ?
• Transport of thousands of macromolecules between
cytoplasm and nucleus occurs through nuclear pore
complex (NPC).
• Nucleocytoplasmic transport is regulated by complex
interplay of nuclear transport machinery which includes
elegant structure like nuclear pore complex ,nuclear
envelope protiens and transport receptors.
5. Why NC transportation is needed ?
• Nuclear envelope separates nucleoplasm from cytoplasm and controls
the nucleocytoplamic transport which is critical for many physiological
and pathological cellular process including gene expression,signal
transduction and oncogenesis.
• RNA transcripts are exported from the nucleus to reach the ribosomes
in the cytoplasm. Proteins such as histones, polymerases, transcription
factors, are imported into nucleus to reach the genetic material.
• Site of DNA replication and RNA biogenesis takes place in the nucleus.
• Site of Protein synthesis is cytoplasm.
6. NUCLEAR ENVELOPE
• Nuclear membrane is a physical barrier b/w nucleoplasm and cytoplasm.It
is a phospholipid bilayer which consists of inner and outer membrane and
are separated by perinuclear space.
• Outer membrane is in continuity with rough endoplasmic reticulum
studded with ribosomes which are engaged in protien synthesis.
• Nuclear membrane is perforated by NPC which fuse the outer nuclear
membrane and inner nuclear membrane together and creates a channel
for nucleocytoplasmic transport.
7. Nuclear Pore Complex
NPC is a multiprotein complex that serve as key regulator of molecular
trafficking b/w cytoplasm and nucleus.
10. CHARACTERISTICS OF NUCLEOCYTOPLASMIC
TRANSPORT
Selectivity:
RNA doesn’t goes through unless accompanied by specific protiens that
act as carriers ,DNA doesnt go through it.
Regulation Of Signals :
Protiens going into the nucleus must have a nuclear localization signal (
NLS) usually a sequence of 4 or more closely located positively charged
amino acids.
Protiens going out of nucleus must have a nuclear export sequence usually
a series of leucine with characteristic spacing b/w them.
12. Mechanism of import and export
Importin + Cargo ( In cytoplasm )→
releases in nucleus on binding with
RanGT
Exportin + Cargo + RanGTP (In
nucleus) → releases on conversion of
RanGTP into RanGDP
Atlanta cook et al
13. IMPORT AND EXPORT OF CARGO
Http7:Annual review in biochemistry/import and export of cargo
14. Ran gtpase cycle
• Ran exists in two conformations depending upon binding of GTP and GDP
• Ran GTPase is very high concentration in the nucleus whereas RanGDP is
very high concentration in cytoplasm.
• Conversion between GTP and GDP is triggered by two Ran specific
regulatory protiens that includes a cytosolic gtpase activating protien GAP
and a nuclear guanine exchange factor GAF.
16. How Directionality Achieved ?
• RanGDP / RanGTP Con.
Gradient
• RanGTP gradient is
maintained by asymmetric
distribution of GAP & GEF
• RanGTP stabilizes export
complex
• RanGDP stabilizes import
complex
SARA NAKIELNY et al
17. Transport Receptor – NTF2
• FG-Nup binding to NTF2
at the spot where Ran
binds upon import
• NTF2 vital for proper
balance of components
across Nuclear envelope
• Nuclear import of
RanGDP is mediated by
NTF2.
• NTF2 spec. recognized by
RanGDP
TIMOTHY et al
18. RNA TRANSPORT
The RNA molecules undergo a variety of post transcriptional processing
events, after which they are transported to their sites of function
throughout the cell.
the subcellular locations of each type of RNA namely messenger RNA
(mRNA), small nuclear RNA (snRNA), ribosomal RNA (rRNA), and transfer
RNA (tRNA) are critical to the normal functioning of the cell.
majority of RNAs are transported to the cytoplasm. Some RNAs need to be
reimported to the nucleus for their function.
All cellular RNA species exit the nucleus through NPCs by an active,
mediated mechanism
19. TRANSPORT OF mRNA
Processing events that influence Nuclear Export
Transcribed pre-mRNAs undergo three co/post transcriptional processing
events.
1. 5’ capping
2. Splicing
3. polyadenylation
20. MECHANISM OF m RNA TRANSPORT
• Trancribed in the nucleus by DNA polymerase II
• Majority of m RNA export is independent of Ran & karyopherin β family.
• The (TAP:p15) m RNA export factor is a heterodimer which helps in Mrna
transport.
21. TRANSPORT OF tRNA
• Transcribed in the nucleus by RNA polymerase Ш.
• t RNA molecules have to be processed before transport into th cytoplam.
• Further modification of tRNA occurs in the cytoplasm.
• Processing comprises
1. Excision of 5’ & 3’ terminal sequences.
2. Addition of CCA to the 3’ end.
3. Base modification.
4. Removal of introns and base editing.
22. RIBOSOMAL RNA TRANSPORT
• Transcribed in the nucleus by RNA polymerase I
• Nuclear export occurs in context to Ribosomal-subunits
• Transport requires energy & through NPC
23. THEMES & PERSPECTIVES
• Common theme :
transport in both
direction across
the NPC mediated
by RNA-binding
proteins
24. FUTURE ISSUES
• In order to determine the general principles of Ran-dependent transport,
the structure of each karyopherin bound cargoes is required.This will
reveal the mechanisms by which nuclear import/export signals,
particularly of cargoes such as NESs and NLS are recognized.
• In some of the cases, the same β-karyopherin can mediate both import
and export so there is need to know extensive studies on karyopherin.
• mRNA export pathway which is one of the complex process and different
mRNAs are recognized for export, process through which it is released to
the cytoplasm hence the nuclear transport machinery performs is need to
be known clearly.
25. Nuclear RNA Export and Its Importance
in Abiotic Stress Responses of Plants
V. Chinnusamy , Z. Gong , and J.-K. Zhu
The transduction of abiotic stresses to switch on genes involved in adaptive responses
are critical to the survival and reproduction of plants exposed to adverse environments.
Plants have evolved multiple stress response pathways, some of which are specific, but
others may be common for various abiotic stresses
Nucleocytoplasmic trafficking—entry of developmental/environmental signals
for gene expression and export of expressed gene products from the nucleus
through nuclear pores—plays a fundamental role in gene expression in eukaryotic
organisms.
26.
27.
28.
29. Role of mRNA Export in Abiotic Stress Response of Plants
• Dead box proteins (DBPs) with RNA helicase activity are involved in RNA metabolism
such as transcription, RNA processing, RNA decay, and nucleocytoplasmic transport. In
yeast and vertebrates, Dbp5 plays a crucial role in export competent Mrnp formation
in nucleus, transport of mRNP through the NPC, and release of mRNA from mRNP in
the cytosol. The role of DBP in mRNA export and abiotic stress response of flowering
plants came from the analysis of the mutant los4 (low expression of osmotically
responsive genes 4) of Arabidopsis .
(GONG et al)
• Role of nups in abiotic stres reaponae was demonstrated ATPNUP 160 mutant which
was hypersensitive to chilling stress .Map based cloning of ATNUP 160 revealed it to
NUP homologus to mammal NUP 160 ATNUP 160 flower show early flowering ATNUP
160 is critical for mrna export , cold resposnive genes expression ,cold tolerance as
well as plant development at normal temperatures
( Dong et al 2006 . Perry et al )
30. SAD 2 domain enhanced gene expression and stress response genes under
cold ABA hypersensitivity in seed germination and seedling growth , SAD 2 is
localized in the nucleus thus result suggested that sad 2 play a crucial role in
nuclear regulator of cold .
(versules et al .2006)
The cold-sensitive phenotype of los4–1 showed inhibited mRNA export
under cold stress conditions. In contrast, the cold-tolerant but heat-sensitive
los4-2 showed normal mRNA export under cold stress but was defective in
mRNA export from the nucleus at warm temperatures (Gong et al. 2005). The
results from los4 mutants demonstrated that LOS4 DEAD-box RNA helicase
plays an important role in mRNA export, which is necessary for abiotic stress
response.
(Gong et al. 2005).
31. CONCLUSION
NUCLEOPHORIN (NUP107–160 complex) proteins shown that they participate
in mRNA export and regulate germination,flowering,hormone response,host-
plant symbiotic interaction, and biotic and abiotic stress tolerance. Further
studies on various components of the nuclear mRNA export machinery in
plants and their role in abiotic stress response will be necessary to better
understanding the link between mRNA export and abiotic stress-responsive
gene expression.
32.
33. EgRBP42 from oil palm enhances adaptation to stress in
Arabidopsis through regulation of nucleocytoplasmic transport of
stress-responsive mRNAs
Wan-Chin Yeap1*, Parameswari Namasivayam2, Tony Eng Keong Ooi1, David Ross Appleton1,
Harikrishna Kulaveerasingam3 and Chai-Ling Ho2, 4*
( Sime Darby Plantation Berhad, Research and Development, Biotechnology and Breeding, Sime
Darby Technology Centre Sdn. Bhd., Serdang, Selangor Darul Ehsan, 43400 Malaysia)
OBJECTIVE - , an hnRNP-like RNA-binding protein from oil palm, which
could be necessary for rapid protein translation to confer abiotic stress
tolerance in plants.
34. MATERIALS AND METHODS
• Six-month-old oil palm seedlings (Dura) were used for abiotic stress
treatments and gene expression analysis.
• Wildtype Arabidopsis, transgenic lines of WT:Pro35SEgRBP42 and mutant
atuba2c were grown under controlled environment conditions at 22 °C, 65
% relative humidity and 120 μmolm-2 s-1 illumination under long day
conditions, 16 h of light followed by 8 h of darkness.
• Pro35S:EgRBP42 in pEG100 and pEG100 (wildtype) were coated on the
surface of gold particles and bombarded into leaf tissue sections, Biolistic
Particle Delivery System bombardments were conducted .
• Total RNA was extracted from oil palm leaves, Arabidopsis leaves and
Arabidopsis flower using NucleoSpin RNA plant (Macherey-Nagel, Düren,
Germany)and observed target gene expression between wildtype .
35.
36. Overexpression of EgRBP42 confers abiotic stress tolerance and accelerates
recovery in vegetative development
45. CONCLUSION
The overexpression of EgRBP42 in Arabidopsis confers
drought, cold, salinity and flood tolerance with enhanced
post-stress recovery. Through this we can propose a model
describing post-transcriptional nucleocytoplasmic transport of
stress-responsive mRNAs in response to different abiotic
stresses, leading to a better adaptation to environmental
changes
Editor's Notes
By contrast to other nuclear envelope proteins, NPC components are evolutionarily conserved between yeast, animals, and plants, indicating that these NPCs share a common progenitor
Individual transport rate of NPC are very high about 500 per sec in both the directions.
Mechanism of Import & Export
Importin binds with cargo in the cytoplasm and releases the cargo molecule in the nucleus on binding with RanGT and Exportin binds with cargo molecule and RanGTP in the nucleus which releases the cargo molecule on conversion of RanGTP into RanGDP. Transport receptors acts as chaperons which shuttle the cargo molecule through the nuclear pore complex. Few examples of transport receptors are Importinβ ,NTF-2 , etc. Ran is responsible for regulating the interaction of transport receptor & cargo. Nucleophorins with specific amino acid sequences involving phenylalanine & glycine is known as FG-Nups (hydrophobic binding site , forms filament & colonizes on both side of NP).
The direction of nucleocytoplsmic transportation of the cargo molecules depend on RanGDP / RanGTP concentration gradient. RanGTP gradient is maintained by asymmetric distribution of GTP activating protein(GAP) & GTP exchange factor (GEF). RanGTP stabilizes export complex and RanGDP stabilizes import complex
Continious movement of ran gtp from the nucleus depletes the nuclear levels of rangtp
Therefore Rangdp has to be reimported into the nucleus and catalyzed
The nuclear import of RanGDP is mediated by NTF2 NTF2 specifically recognizes Ran in its GDP-bound state
The continuous outflow of exportin-cargo complexes and
cargo-free importins from the nucleus would lead to depletion
of nuclear RanGTP and eventual shutdown of all
nuclear transport, thus necessitating that Ran be restored
to the nucleus. The return of Ran is mediated by another
transport factor, NTF2 [34, 35]. NTF2 is unrelated to the
importin b family, and it binds only RanGDP. Upon entering
the nucleus, NTF2 and RanGDP must dissociate
from each other, and Ran’s GDP must be replaced with
GTP. Both steps are fostered by a Ran-specific guanine
nucle
Most eukaryotic RNAs are produced in the nucleus by RNA polymerase I, II, or III.
Processing Events That Influence Nuclear Export: Most mRNAs are transcribed as pre-mRNAs, which generally undergo three posttranscriptional processing events: 50 capping, i.e. addition of a 7-monomethyl guanosine (m7G) cap structure to the 50 end of the transcript; removal of introns by splicing; and polyadenylation at a defined site within the 30 untranslated region
RIBOSOMAL RNA TRANSPORT: It is a common precursor that is completely processed and assembled into ribosomal subunits, all within the nucleolus Therefore, nuclear export of these rRNAs occurs in the context of ribosomal subunits. Beyond the fact that ribosomal subunit export is a unidirectional, saturable process that requires energy and All rRNAs except 5S rRNA are transcribed occurs through NPCs very little is known about export of ribosomes.
Relatively little information exists on the NUPs and NEFs required for mRNA export in plants
Bioinformatics analyses have identified several conserved NUPs from rice and Arabidopsis that are homologous to yeast and vertebrate NUPs (Neumann et al.2006). Since the nuclear mRNA export pathway is highly conserved across species (fungi, insects, vertebrate) and many proteins similar to NUPs and NEFs have been found in higher plants, the mRNA export process in higher plants appears to be similar to that of other eukaryotes.