Cloning, Expression Profile, and Genomic Organization of the Mouse STAP/A170 Gene

• Published in: Genomics (San Diego, Calif.) Calif.), 1999-08, Vol.60 (1), p.87-95
• Main Authors: Okazaki, Makoto, Ito, Sachiko, Kawakita, Koji, Takeshita, Sunao, Kawai, Shinji, Makishima, Fusao, Oda, Hiroaki, Kakinuma, Atsushi
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Cloning, Molecular; DNA - chemistry; DNA - genetics; DNA, Complementary - chemistry; DNA, Complementary - genetics; Exons; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes - genetics; Genes. Genome; Heat-Shock Proteins - genetics; Introns; Male; Mice; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Regulatory Sequences, Nucleic Acid; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Sequestosome-1 Protein; TATA Box; Tissue Distribution; Transcription, Genetic
• ISSN: 0888-7543; 1089-8646
• DOI: 10.1006/geno.1999.5902

The preferential screening of cDNA libraries derived from the mouse osteoblastic cell line MC3T3-E1 has yielded a cDNA clone encoding a 442-amino-acid protein designated STAP (signal transduction and adaptor protein), which contains several motifs shared among transcription factors and adaptors such as a Zn-finger like motif, a proline-rich domain, and a PEST sequence. The amino acid sequence homology search also reveals that STAP is identical to a mouse oxidative stress protein, A170, and has 90% homology with a human p62 protein that binds to the tyrosine kinase p56lck SH2 domain. Northern blot analysis indicated a broad expression profile of STAP mRNA in various tissues and cell lines. In MC3T3-E1 cells, STAP mRNA was induced by treatment with TGF-β, but not with BMP-2 or GDF-5. Analysis of the mouse STAP gene isolated from the genomic library revealed that the STAP gene spans a region of over 11 kb and comprises eight exons. The transcription start site was identified by primer extension analysis to be located 35 bp upstream from the translation initiation site. Sequencing analysis of the 5′ flanking region of the STAP gene revealed multiple consensus motifs/sequences for several DNA binding transcription factors. The STAP gene had a TATA box, but no CCAAT box. Potential Sp1, AP-1, NF-E2, MyoD, and NF-κB binding sites were found in the 5′ flanking region (1.4 kb) of the STAP gene.