Characterization of the human PRIP-1 gene structure and transcriptional regulation

Ayako Murakami, Miho Matsuda, Akihiko Nakasima, Masato Hirata

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The PRIP [phospholipase C related, but catalytically inactive protein] family has been isolated as a novel inositol 1,4,5-trisphosphate binding protein with a domain organization similar to phospholipase C-δ but lacking the enzyme activity, comprising PRIP-1 and PRIP-2. The PRIP-1 gene is expressed predominantly in the brain, while PRIP-2 exhibits a relatively ubiquitous expression in rats and mice. We also found that PRIP-1 plays an important role in type A receptor signaling for γ-aminobutyric acid in the brain. In this study, we investigated PRIP-1 gene structure and the possible mechanisms involved in the expression. The tissue distribution pattern of PRIP gene expression in humans was similar to that in rodents. 5′RACE (rapid amplification of cDNA ends) analysis using PRIP-1 gene specific primers with human brain mRNA revealed the presence of three new exons, indicating that the PRIP-1 gene is organized into 8 exons intervened by 7 introns. Although three transcripts resulting from the alternative splicing of exon 2 and/or 3 were detected, a transcript lacking exons 2 and 3 was predominantly expressed in humans, suggesting that the translation start codon of human PRIP-1 exists in exon 1. To characterize the human PRIP-1 promoter, transient luciferase assay was carried out with luciferase constructs including various lengths of the 5′ flanking region of the PRIP-1 gene. The results indicated that the positive regulatory region is located - 237 to - 108 bp upstream from the transcription start site. Gel shift assay revealed the specific binding of some nuclear proteins to this region, suggesting that the existence of transcription factors contributes to the positive regulation of PRIP-1 gene expression. Mutation analyses revealed that the binding of a transcription factor, MAZ to the regulatory site leads to the promoter activity, indicating that MAZ is involved in the expression regulation of the human PRIP-1 gene.

Original languageEnglish
Pages (from-to)129-139
Number of pages11
JournalGene
Volume382
DOIs
Publication statusPublished - Nov 1 2006

Fingerprint

Exons
Genes
Type C Phospholipases
Luciferases
Brain
Transcription Factors
Aminobutyrates
Gene Expression
Inositol 1,4,5-Trisphosphate
Initiator Codon
5' Flanking Region
Nucleic Acid Regulatory Sequences
Transcription Initiation Site
Alternative Splicing
Tissue Distribution
Nuclear Proteins
Rodentia
Carrier Proteins
Complementary DNA
Gels

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Characterization of the human PRIP-1 gene structure and transcriptional regulation. / Murakami, Ayako; Matsuda, Miho; Nakasima, Akihiko; Hirata, Masato.

In: Gene, Vol. 382, 01.11.2006, p. 129-139.

Research output: Contribution to journalArticle

Murakami, Ayako ; Matsuda, Miho ; Nakasima, Akihiko ; Hirata, Masato. / Characterization of the human PRIP-1 gene structure and transcriptional regulation. In: Gene. 2006 ; Vol. 382. pp. 129-139.
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