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Genomic organization of mouse gene zfp162
C Wrehlke1, W R Wiedemeyer, H P Schmitt-Wrede
1Division of Molecular Parasitology and Centre for Biological and Medical Research, Heinrich-Heine-University, Düsseldorf, Germany.
Abstract:
We report the cloning and characterization of the alternatively spliced mouse gene zfp162, formerly termed mzfm, the homolog of the human ZFM1 gene encoding the splicing factor SF1 and a putative signal transduction and activation of RNA (STAR) protein. The zfp162 gene is about 14 kb long and consists of 14 exons and 13 introns. Comparison of zfp162 with the genomic sequences of ZFM1/SF1 revealed that the exon-intron structure and exon sequences are well conserved between the genes, whereas the introns differ in length and sequence composition. Using fluorescent in situ hybridization, the zfp162 gene was assigned to chromosome 19, region B. Screening of a genomic library integrated in lambda DASH II resulted in the identification of the 5'-flanking region of zfp162. Sequence analysis of this region showed that zfp162 is a TATA-less gene containing an initiator control element and two CCAAT boxes. The promoter exhibits the following motifs: AP-2, CRE, Ets, GRE, HNF5, MRE, SP-1, TRE, TCF1, and PU.1. The core promoter, from position -331 to -157, contains the motifs CRE, SP-1, MRE, and AP-2, as determined in transfected CHO-K1 cells and IC-21 cells by reporter gene assay using a secreted form of human placental alkaline phosphatase. The occurrence of PU.1/GRE supports the view that the zfp162 gene encodes a protein involved not only in nuclear RNA metabolism, as the human ZFM1/SF1, but also in as yet unknown macrophage-inherent functions.
Insights
We cloned and characterized the mouse zfp162 gene, a homolog of human SF1. This gene is involved in RNA metabolism and potentially macrophage functions.
Area of Science:
- Molecular Biology
- Genetics
Background:
- The mouse gene zfp162, previously known as mzfm, is the homolog of the human ZFM1 gene.
- ZFM1 encodes the splicing factor SF1 and a putative signal transduction and activation of RNA (STAR) protein.
Purpose of the Study:
- To clone and characterize the alternatively spliced mouse gene zfp162.
- To investigate the genomic structure, chromosomal localization, and promoter elements of zfp162.
Main Methods:
- Gene cloning and characterization.
- Fluorescent in situ hybridization for gene mapping.
- Genomic library screening and sequence analysis.
- Reporter gene assays in transfected cells.
Main Results:
- The zfp162 gene is approximately 14 kb long, with 14 exons and 13 introns, showing conserved exon-intron structure with human ZFM1/SF1.
- zfp162 was mapped to mouse chromosome 19, region B.
- Sequence analysis revealed a TATA-less promoter with initiator control elements, CCAAT boxes, and multiple transcription factor binding motifs (AP-2, CRE, Ets, GRE, HNF5, MRE, SP-1, TRE, TCF1, PU.1).
- Reporter gene assays confirmed the activity of the core promoter (-331 to -157) containing CRE, SP-1, MRE, and AP-2 motifs.
Conclusions:
- The mouse zfp162 gene is structurally conserved with its human homologue, ZFM1/SF1.
- The promoter analysis suggests roles in nuclear RNA metabolism and potentially unique macrophage functions due to the presence of PU.1/GRE motifs.