Rhoads D D, Dixit A, Roufa D J
Mol Cell Biol. 1986 Aug;6(8):2774-83. doi: 10.1128/mcb.6.8.2774-2783.1986.
Chinese hamster ribosomal protein S14 cDNA was used to recognize homologous human cDNA and genomic clones. Human and Chinese hamster S14 protein sequences deduced from the cDNAs are identical. Two overlapping human genomic S14 DNA clones were isolated from a Charon 28 placental DNA library. A fragment of single-copy DNA derived from an intron region of one clone was mapped to the functional RPS14 locus on human chromosome 5q by using a panel of human X Chinese hamster hybrid cell DNAs. The human S14 gene consists of five exons and four introns spanning 5.9 kilobase pairs of DNA. Polyadenylated S14 transcripts purified from HeLa cell cytoplasma display heterogeneous 5' ends that map within noncoding RPS14 exon 1. This precludes assignment of a unique 5' boundary of RPS14 transcripts with respect to the cloned human genomic DNA. Apparently HeLa cells either initiate transcription at multiple sites within RPS14 exon 1, or capped 5' oligonucleotides are removed from most S14 mRNAs posttranscription. In contrast to the few murine ribosomal protein and several other mammalian housekeeping genes whose structures are known, human RPS14 contains a TATA sequence (TATACTT) upstream from exon 1. Three related short sequence motifs, also observed in murine and yeast ribosomal protein genes, occur in this region of the RPS14 gene. RPS14 introns 3 and 4 both contain Alu sequences. Interestingly, the Alu sequence in intron 3 is located slightly downstream from a chromosome 5 deletion breakpoint in one human X hamster hybrid clone analyzed.
用中国仓鼠核糖体蛋白S14 cDNA识别同源的人类cDNA和基因组克隆。从cDNA推导的人类和中国仓鼠S14蛋白序列是相同的。从Charon 28胎盘DNA文库中分离出两个重叠的人类基因组S14 DNA克隆。通过使用一组人类×中国仓鼠杂交细胞DNA,将来自一个克隆的内含子区域的单拷贝DNA片段定位到人类染色体5q上的功能性RPS14基因座。人类S14基因由5个外显子和4个内含子组成,跨越5.9千碱基对的DNA。从HeLa细胞细胞质中纯化的聚腺苷酸化S14转录本显示出异质的5'末端,这些末端定位在非编码RPS14外显子1内。这排除了根据克隆的人类基因组DNA确定RPS14转录本独特的5'边界。显然,HeLa细胞要么在RPS14外显子1内的多个位点起始转录,要么在转录后从大多数S14 mRNA上去除了加帽的5'寡核苷酸。与少数已知结构的小鼠核糖体蛋白和其他几个哺乳动物管家基因不同,人类RPS14在外显子1上游含有一个TATA序列(TATACTT)。在RPS14基因的这个区域还出现了三个相关的短序列基序,在小鼠和酵母核糖体蛋白基因中也观察到。RPS14内含子3和4都含有Alu序列。有趣的是,在分析的一个人类×仓鼠杂交克隆中,内含子3中的Alu序列位于5号染色体缺失断点的稍下游。