| dc.creator |
Butta, Nora |
|
| dc.creator |
Larrucea, Susana |
|
| dc.creator |
Alonso, Sonia |
|
| dc.creator |
Rodríguez, Ramón B. |
|
| dc.creator |
García Arias-Salgado, Elena |
|
| dc.creator |
Sánchez Ayuso, Matilde |
|
| dc.creator |
González-Manchón, Consuelo |
|
| dc.creator |
Parrilla, Roberto L. |
|
| dc.date |
2008-04-07T09:13:20Z |
|
| dc.date |
2008-04-07T09:13:20Z |
|
| dc.date |
2006-05-09 |
|
| dc.date.accessioned |
2017-01-31T01:01:35Z |
|
| dc.date.available |
2017-01-31T01:01:35Z |
|
| dc.identifier |
BMC Mol Biol.:7:17(2006) |
|
| dc.identifier |
1471-2199 |
|
| dc.identifier |
http://hdl.handle.net/10261/3452 |
|
| dc.identifier |
10.1186/1471-2199-7-17 |
|
| dc.identifier.uri |
http://dspace.mediu.edu.my:8181/xmlui/handle/10261/3452 |
|
| dc.description |
12 pages |
|
| dc.description |
[Background] Podocalyxin (podxl) is a heavily glycosylated transmembrane protein mainly found
on the apical membrane of rat podocytes and also in endothelial, hematopoietic, and tumor cells.
Despite of its interest no much is known about the transcriptional regulation of podxl in different
cells. Thus, we aimed at studying the functional features of the 5'-regulatory region of the human
Podxl gene. |
|
| dc.description |
[Results] The promoter region of the human Podxl gene has been cloned and its structure and
function were analyzed. The primary DNA sequence is rich in G+C and is devoid of TATA or
CAAT boxes. The sequence contains recognition sites for several putative transcription factors;
however, the basic promoter activity seems to rely entirely on Sp1 transcription factor since
supershift analysis was positive only for this factor. The region encompassed by 66 to -111 nts
conferred the minimal transcriptional activity that increases as the number of Sp1 sites augmented
with the length of the promoter fragment. In Sp1-lacking insect cells the Podxl promoter constructs
showed activity only if cotransfected with an Sp1 expression plasmid. Finally, mutation of the Sp1
sites reduced the promoter activity. We analyzed whether methylation of the CpG dinucleotides present in the first ~600 nts of the
promoter region of Podxl could explain the variable rates of expression in different types of cells.
Inactivation of methyltransferases by 5'-aza-2'deoxicitidine showed a dose-dependent increase in
the podxl content. Moreover, in vitro methylation of the promoter constructs -111,-181 and -210
led to an almost complete reduction of the promoter activity. A correlation was found between
the degree of methylation of the CpG promoter dinucleotides and the rate of podxl expression in
different cell lines. |
|
| dc.description |
[Conclusion] Our results indicate that transcriptional regulation of Podxl is supported primarily by
Sp1 site(s) and that DNA-methylation of the CpG promoter islands contributes to control the
tissue specific expression of podxl. |
|
| dc.description |
N. Butta is recipient of a tenure track grant Ramon y Cajal from the Spanish
Ministry of Science. S. Larrucea was supported by a contract from the
Comunidad de Madrid (08.4/0015.1/2001) and at present holds a research
tenure track of the Spanish Council of Research. S. Alonso was recipient of
a predoctoral fellowship from the Basque Autonomous Community
(BF101-40). E G Arias-Salgado is recipient of a tenure research grant Juan
de la Cierva. This work has been supported in part by grants from the
Direccion General de Investigacion (SAP 2004-04345), Fondo de Investigaciones
Sanitarias (FIS-PI021263 and PI050546) and Comunidad de Madrid
(08.4/0029.1/2003). N. Butta was recipient of a grant from the Fundacion
Rodríguez Pascual. We thank Dr. Kershaw for the generous gift of the mAb
3D3. |
|
| dc.description |
Peer reviewed |
|
| dc.format |
487082 bytes |
|
| dc.format |
application/pdf |
|
| dc.language |
eng |
|
| dc.publisher |
BioMed Central |
|
| dc.relation |
Publisher’s version |
|
| dc.relation |
http://www.biomedcentral.com/1471-2199/7/17 |
|
| dc.rights |
openAccess |
|
| dc.title |
Role of transcription factor Sp1 and CpG methylation on the regulation of the human podocalyxin gene promoter |
|
| dc.type |
Artículo |
|