Functional characterization of the ribonucleoprotein, PTB-binding 1/Raver1 promoter region.
Raver1 is a ribonucleoprotein, evolutionarily conserved in mammals, which acts as a polypyrimidine tract-binding protein (PTB/hnRNPI) co-repressor in regulating alternative splicing events. The mouse homologue has been identified as a dual compartment protein that interacts with PTB within perinucleolar structures, and localizes at microfilament plasma membrane attachment sites in fibroblasts, epithelial and muscle cells. Human Raver1 gene is localized on chromosome 19p13.2 and encodes for an inferred 756 amino acid protein sharing 87% similarity with the mouse orthologue. The human Raver1 gene expression has not been previously investigated. Here we report the mRNA expression profile of human Raver1 gene and the molecular characterization of its promoter region. From the in silico analysis of 1542 bp of the Raver1 5'-flanking region (GC content=61%), no canonical TATA or CAAT boxes can be highlighted, whereas several consensus Sp1 putative binding sequences can be predicted within 1 kb from the transcription start site (TSS) that we determined by 5'-RACE. Functional analyses established a minimal region involved in the regulation of the human Raver1 promoter activity. Mutational analyses and transfection studies indicated that a GGGAGCTCCC sequence at -531 represents a putative cis signal acting as a negative regulator element of the promoter function. Altogether, our results indicate that human Raver1 gene promoter region shares common features of ubiquitously expressed gene with the interacting splicing regulator PTB.
Romanelli
Maria Grazia
MG
Department of Mother and Child, Biology and Genetics, University of Verona, Strada le Grazie 8, 37134 Verona, Italy. mariagrazia.romanelli@univr.it
Lorenzi
Pamela
P
Avesani
Francesca
F
Morandi
Carlo
C
D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000502:physiology*; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D004247:DNA; D017931:DNA Primers; D018076:DNA, Complementary; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D009687:Nuclear Proteins; C486705:RAVER1 protein, human; D012261:Ribonucleoproteins; D004247:DNA