| RRC ID |
19171
|
| Author |
Tominaga A, Sugawara H, Futagawa T, Inoue K, Sasaki K, Minamino N, Hatakeyama M, Handa H, Miyata A.
|
| Title |
Characterization of the testis-specific promoter region in the human pituitary adenylate cyclase-activating polypeptide (PACAP) gene.
|
| Journal |
Genes Cells
|
| Abstract |
Pituitary adenylate cyclase-activating polypeptide (PACAP) is a pleiotropic neuropeptide localized in the testis at concentration comparable to that found in the brain, suggesting involvement in spermatogenesis. In this study, we identified the human PACAP testis-specific exon (TSE) 10.9 kb upstream from the translational start site and found that the testis-specific transcript of the human PACAP gene was found to be spliced from the TSE into a region of intron 2 without a frameshift. The resulting PACAP precursor has no signal peptide, suggesting that PACAP functions physiologically in an intracrine manner in the testis. The 5'-flanking region of the TSE contains an 80-bp fragment with potent promoter activity in testicular F9 cell. Electrophoresis mobility shift assays showed that proteins from the F9 nuclear extract interacted specifically with the 80-bp fragment. DNA affinity chromatography allowed isolation of the specific proteins bound to the 80-bp fragment, two of which were identified as Poly (ADP-ribose) polymerase-1 (PARP-1) and TIA-1-related protein (TIAR) by mass spectrometry. By using their siRNAs, the depletion of their proteins in F9 cells affected the potent promoter activity of the 80-bp fragment, suggesting that they might be involved in the testis-specific gene expression of PACAP.
|
| Volume |
15(6)
|
| Pages |
595-606
|
| Published |
2010-6-1
|
| DOI |
10.1111/j.1365-2443.2010.01403.x
|
| PII |
GTC1403
|
| PMID |
20500521
|
| MeSH |
Alternative Splicing
Amino Acid Sequence
Animals
Base Sequence
Binding Sites / genetics
Cell Line, Tumor
Electrophoretic Mobility Shift Assay
Exons / genetics
Gene Expression Profiling*
Humans
Luciferases / genetics
Luciferases / metabolism
Male
Mice
Molecular Sequence Data
Pituitary Adenylate Cyclase-Activating Polypeptide / genetics*
Pituitary Adenylate Cyclase-Activating Polypeptide / metabolism
Poly(ADP-ribose) Polymerases / metabolism
Promoter Regions, Genetic / genetics*
Protein Binding
RNA Interference
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Swiss 3T3 Cells
Testis / metabolism*
Transfection
|
| IF |
1.655
|
| Times Cited |
7
|
|
WOS Category
|
GENETICS & HEREDITY
CELL BIOLOGY
|
| Resource |
| Human and Animal Cells |
NEC14(RCB0490) |
