Identification and analysis of the human neural polypyrimidine tract binding protein (nPTB) gene promoter region.
Neural polypyrimidine tract-binding protein nPTB, originally identified as the neuronal counterpart of the hnRNPI/PTB protein, is an RNA binding protein involved into tissue-specific alternative splicing regulation. Here we describe the functional characterization of the promoter sequence of nPTB in HeLa and neuroblastoma cells. By means of genomic sequence analysis we have isolated and cloned a 1587-base pair region upstream the human nPTB coding region. The nPTB proximal promoter, although rich in G+C content and presenting putative binding sites for the transcription factors Sp1, NF-1, NF-kB and Oct-1, lacks a typical TATA box. Luciferase transient expression assays using deletion mutants have identified the proximal promoter region at -125 relative to the transcription start site. Alignment of human, murine and chimpanzee genomic sequences upstream the nPTB exon 1 has provided evidences for the evolutionary conservation of specific transcription factor binding sites.
Department of Mother and Child, Biology and Genetics, University of Verona, Strada le Grazie 8, 37134 Verona, Italy. mariagrazia.romanelli@univr.it
Maria Grazia
MG
Romanelli
Pamela
P
Lorenzi
Carlo
C
Morandi
D024506:5' Flanking Region / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D051839:NFI Transcription Factors; D009419:Nerve Tissue Proteins / Q000235:genetics*; D050811:Octamer Transcription Factor-1; D038941:Polypyrimidine Tract-Binding Protein / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment / Q000379:methods; D012689:Sequence Homology, Nucleic Acid; D016329:Sp1 Transcription Factor / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D024363:Transcription Initiation Site; D014162:Transfection
D004268:DNA-Binding Proteins; D016328:NF-kappa B; D051839:NFI Transcription Factors; D009419:Nerve Tissue Proteins; D050811:Octamer Transcription Factor-1; C494165:POU2F1 protein, human; C504334:PTBP2 protein, human; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; C080333:transcription factor nuclear factor 1; D038941:Polypyrimidine Tract-Binding Protein; D008156:Luciferases