Roberts S G, Layfield R, Bannister A J, McDonald C J
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, England.
Eur J Biochem. 1991 Dec 18;202(3):969-74. doi: 10.1111/j.1432-1033.1991.tb16457.x.
A mouse genomic B-type proline-rich protein (PRP) cosmid clone was isolated by cDNA hybridisation and mapped, the gene region was subcloned and 3770 bp were sequenced. This gene (MP4) contained three introns and encoded a 1020-nt (nt, nucleotide) mRNA for a PRP precursor 300 amino acids long arranged with 11 imperfect 18-residue proline-rich repeats. The transcriptional start point was determined by S1 nuclease mapping and primer extension to be 26 bp downstream of a TATAA sequence. Sequence comparisons revealed that only two regions from positions -650 bp - -30 bp were highly conserved in all other PRP genes, PRP boxes 1 and 2. Box 1 at positions -112 to -135 contained ets-like and rel/NFkB-like elements and was 74% conserved over 23 bp. Box 2 at positions -33 - -51 was 53% conserved over 19 bp. A search of the EMBL and GenBank sequence libraries indicated that PRP box 1 was only present upstream of the known mammalian PRP gene sequences and was absent from other genes. These conserved sequences may thus be relevant to the tissue-specific and beta-adrenergic regulation of PRP gene transcription.
通过cDNA杂交分离并定位了一个小鼠基因组B型富含脯氨酸蛋白(PRP)黏粒克隆,对该基因区域进行亚克隆并测序了3770 bp。该基因(MP4)包含三个内含子,编码一个1020 nt(nt,核苷酸)的mRNA,用于一个300个氨基酸长的PRP前体,其排列有11个不完美的18个残基的富含脯氨酸重复序列。通过S1核酸酶图谱分析和引物延伸确定转录起始点位于TATAA序列下游26 bp处。序列比较显示,在所有其他PRP基因中,只有从-650 bp至-30 bp的两个区域高度保守,即PRP框1和框2。位于-112至-135的框1包含ets样和rel/NFkB样元件,在23 bp上74%保守。位于-33至-51的框2在19 bp上53%保守。对EMBL和GenBank序列文库的搜索表明,PRP框1仅存在于已知哺乳动物PRP基因序列的上游,其他基因中不存在。因此,这些保守序列可能与PRP基因转录的组织特异性和β-肾上腺素能调节有关。