Realizado por: Salomé Rivera Martinez y Juliana Vargas Arboleda

1. SALOMÉ RIVERA MARTINEZ
JULIANA VARGAS ARBOLEDA
Tercer semestre
2017-1

2. INTRODUCTION
ELLIPTOCYTOSIS
Also known as
ovalocytosis, is an
inherited disorders
where the erythrocytes
are elliptical rather
than the typical
biconcave disc shape.

3. INTRODUCTION
X-LINKED RECESSIVE SYNDROME

4. INTRODUCTION
AMMECR1
Is a protein encoded by the
AMMECR1 gene on human
chromosome Xq22.3. Is localized
in the critical region of contiguous
deletion syndrome implicated in
Alport syndrome, mental
retardation, midface hypoplasia,
and elliptocytosis chromosomal
region gene 1.

5. INTRODUCTION
RELATION
A truncating mutation in AMME chromosomal
region gene 1 (AMMECR1) gene is leading
candidate due to X-linked inheritance that is
characterized by the presence of elliptocytosis
and midface hypoplasia in the proband.

6. OBJETIVE
Provide further evidence that
mutated AMMECR1 gene is
responsible for the clinically
recognizable X-linked condition
with variable expressivity.

7. MÉTODOS
1. Sujetos: Niño de 4 años nacido de padres no
consanguíneos y no afectados, de origen judío , libio,
yemenita y turco; hermana no afectada y tio materno
afectado.

8. MÉTODOS

9. MÉTODOS
2. Secuenciación:
Extracción de DNA Fragmentación
por sonicación
Centrifugación
PurificaciónLimpieza
Procesamiento
de muestras
alineación del genoma y
selección de variantes
Lectura

10. 2. Secuenciación:
Utilización del método enzimático o de Sanger,
en el cual no se degrada el DNA sino que se
interrumpe de forma controlada de la síntesis
de una hebra complementaria.
Forward: 5'- cccgtggacatttggtcattgg-3 ', Reverse 5'-ggcacctaacctagcagacagtcaatactc-3'.
MÉTODOS

11. MÉTODOS
3. PCR
➝Se extrajo RNA de la sangre periférica de el
niño y la madre.
➝ Se sintetizó ADN complementario y se
amplifico el AMMERC1 con RT PCR:
Forward: 5'-GATGGTGGTGTCAGCAGAGAT -3'.
Reverse: 5'- CAGGAATAATGGTTGTATGGCGG -
3'.
➝ se cargó y pasó por un gel de agarosa al 1.5%.

12. MÉTODOS

13. RESULTADOS

14. RESULTADOS
Fig. 3. Alternative splicing ofAMMECR1. (A)AMMECR1 schematic representation of exons, numbered 1 to 6, in theWild-type(WT) isoforms
and Patient (II-2) isoforms. Themutation in the patient transcript is indicated on alternative exon 2.
(B) PCR amplification and separation in Agarose gel of AMMECR1 from proband and mother. Upper bands represent the exon 2
inclusion events, while the lower represent the exon 2 skipping events. Bands were excised, extracted and sequenced for confirmation.

15. RESULTADOS
Fig. 3. (C) AMMECR1 isoforms (taken from UCSC Genome Browser website; https://genome.ucsc.edu/) show that in one out of the three
isoforms exon 2 is skipped (missing box, indicated by arrow). The horizontal line represents the introns while the vertical boxed indicate the
exons. The image is drawn to scale. Note that the bottom isoform starts further upstream to this current view.
(D) PCR amplification as in B on eight healthy individuals.

16. RESULTADOS
Fig. 4. AMMECR1 protein model demonstrating the structural importance of position R45 on exon 2 of the protein. (A) AMMECR1 protein ribbon is colored by conservation (maroonwhite-
cyan scale from most conserved to least conserved positions) and R45 is shown in CPK representation, with the nitrogen atoms in blue and the carbons in conserve colors. R45
is located on a loop between two beta sheets that interact with an alpha helix on the core of a subdomain. (B) R45 is close to a 6-amino-acid motif (LRGCIG), colored in maroon, that is
highly conserved throughout evolution and is thus probably functionally important. (C) A change on a nearby loop, or skipping of this region which is located on exon 2 of the gene,
will lead to protein complete destabilization and probably lack of this entire protein. (For interpretation of the references to color in this figure legend, the reader is referred to the
web version of this article.)

17. DISCUSSION
ACTOR WHAT SAID YES OR NOT
Vitelli et al AMMECR1 gene encodes a
protein with a nuclear
localization and presently
unknown function, yet was
suggested to play a role as a
regulatory protein.
Meloni et al FACL4 gene has been
implicated in intellectual
disability in patients with
deletions including this gene.

18. DISCUSSION
ACTOR WHAT SAID YES OR NOT
Andreoletti et al Clinical features shared by the
individuals escribed in this report and
individuals described by Andreoletti et
al. include cleft palate, club feet, a flat
facial profile with a flat nasal bridge,
thin lips micrognathia,hearing loss and
eliptocytosis.
Balaji and Aravind AMMECR1 has previously been linked
to involvement in an uncharacterized
RNA base modification, potentially
entailing the transfer of a modifying
group onto an RNA base via a
conserved cysteine.

19. CONCLUSION
• It is a dissease that can be treated but not cured,
since we can improve the patient's lifestyle but not
prevent the gene from expressing itself.
• This disease is little previous and the largest number
of cases have been recorded in central and western
Africa.
• With the RT PCR the expression of the mutated gene
in the proband could be determined.
• The clinical characteristics of patients with
AMMECR1 gene mutation are not always the same.

20. Juliana Vargas Arboleda