1. Einat zalckvar, Christina Paulus, Desiree Tillo, Alexandra Absach-Nitzsche, Yaniv Lubling, Carla
Winterling, Nicholas Strieder, Katrin Mucke, Felicia Goodrum, Eran Segal, Michael Nevels.
Sara Puche Gómez
Andrés Felipe Restrepo Ramírez
Medicine students
III Semester
Molecular Biology
UPB

2. Associated diseases:
• Infectious mononucleosis
• Congenital Infection  Cytomegalic inclusion disease
• Pneumonitis
• Gastrointestinal disease

3. Chromatin
Complex
between
eukaryotic
DNA and
proteins
Constitutes
chromosome
Have histone
and non-
histone
proteins
Basic
structure is
the
nucleosome

4. 146 pairs of
DNA coiled
1,65 times
Histones H2A,
H2B, H3 AND
H4, and them
with H1 form
chromatosome
Nucleosome

5. Its the first set of viral gene
products to be expressed
Not require the novo protein
synthesis for their expression
IE 2: essential transcriptional
transactivator. Genes activated
and inhibits downstream IE gene
transcription through their
specific binding to DNA
IE 1: promiscuous transactivator.
Inactive histone deacetylase
(HDAC) to repress gene
expression can viral
IE Protein

6. Cytomegalovirus
Have IE
protein
Which
expresses to
help
The virus to
enter the cell
nucleus
And use
proteins
To form
nucleosomes
And then
chromatin
So then is
incorporated
in
DNA

7. To investigate the role of the human cytomegalovirus IE
protein in the temporal organization of the nucleosome and
involvement in the function and structure of chromatin

8. PCR
•No es una tecnica analìtica, es una metología
•Desnaturalización del DNA para dar dar hebras sencillas
• Hibridación específica de la hebra sencilla con un primer
•Replicación de la hebra sencilla por una DNA polimerasa a partir del primer anterior
PCRq
•Basada en el analisis digital de imagen mediante cualquier programa de analisis de
imagen, permite realizar una precisa cuantificación de bandas de ácidos nucléicos
desarrollados en gel de agarosa
PCR real
time
•Mas seguro que la PCR, procesos como la amplificación y la detección se dan
simultaneamente y por fluorescencia se puede detectar la producción de DNA en el
momento

10. RESULTS

11. FIGURA 1
cambios masivos en la organización del nucleosoma en todo el
genoma hCMV durante la infección productiva.

12. FIGURA 1

13. FIGURA 2
Cambios temporales IE1-dependientes en la organización del
nucleosoma en el genoma del hCMV

14. FIGURA 2

15. FIGURA 3
Efectos IE1-dependientes en la ocupación
nucleosomal a través del hCMV genoma

16. FIGURA 3

17. FIGURA 4
Correlación
negativa entre
cambios IE1
dependientes en
la organización
y expresión del
nucleosoma en
el genoma del
hCVM

18. DISCUSIÓN

19. REFERENCES WHAT HE OR SHE SAY? AGREE/DISAGREE
Nevels M et al
herpesvirus
genomes are
commonly
considered to form
regular nucleosomal
arrays resembling
bulk cellular
chromatin in
Latently infected
cells
¿Why?

20. This type of chromatin organization has
not been formally shown for hCMV or
any β-herpesvirus.
1
Herpesvirus histones inside virus particles
Nonnucleosomal structure of viral genomes
nuclei of cells infected with herpes viruses
2

21. REFERENCES WHAT HE OR SHE
SAID?
AGREE / DISAGREE
Greaves RF et al
Disruption of IE1
delays
transcription
during productive
hCMV infection
AGREE

22. REFERENCES
WHAT HE OR SHE
SAID?
AGREE/DISAGREE
Sekinger et al
Genome-wide
analyses in yeast
have indicated that
nucleosome
occupancy
correlates poorly
with transcription
It was found significant negative correlations between
changes in nucleosome occupancy and changes in gene
expression between WT and dlIE1 strains 96 h postinfection.

23. REFERENCES WHAT HE OR SHE
SAY?
AGREE/DISAGREE
Munch K et al
IE1 is not
considered to bind
DNA directly , the
viral protein
associates with
chromatin and
nucleosome
modifying factors
IE1 on nucleosome occupancy
and organization might be mediated through direct
interaction
between the viral protein and chromatin components
AGREE

24. conclusions

25.  Increasingly more importance is been given to the complex role of
proteins in the chromatin organization and gene expression
 understand the mechanism during productive hCMV infection
would help to develop new treatments for patients worldwide.
 In order to properly understand and analyze a research article is
necessary to study deeply the experimental models and the basis of
the used techniques to understand the housed results, and the
choice of appropriate techniques is essential to get accurate results
consistent with the investigation purpose.
 The Chip on chip technique is needed to confirm the results because
it show the interaction between DNA and nuclear proteins as
histones

26. MAPAS

27. Sara Puche
GSara