Ozer J, Chalkley R, Sealy L
Department of Cell Biology, Vanderbilt University Medical Center, Nashville, TN 37232.
Gene. 1993 Feb 28;124(2):223-30. doi: 10.1016/0378-1119(93)90397-l.
The genomic copy multiplicity of the CCAAT transcription complex component enhancer factor I subunit A (EFIA) has been examined. When a mammalian genomic Southern blot was hybridized to a rat EFIA cDNA, a complex pattern consisting of numerous related sequences was found in all the species examined, with Bos taurus being the least complex. An EFIA#1 cDNA from Bos taurus was isolated from a primary lung endothelial cell cDNA library by screening with the 1489-bp rat EFIA cDNA. The deduced bovine EFIA#1 amino acid (aa) sequence is 98% identical to rat EFIA and 100% identical to human EFIA/DbpB/YB-1 family member DNA-binding protein B (DbpB). In addition, a processed EFIA pseudogene from Bos taurus, designated bovine psi EFIA#1, was obtained from a genomic library by screening with a rat EFIA cDNA probe. The bovine psi EFIA#1 gene has an ORF which, if expressed, would encode a 140-aa sequence, with aa 31-140 having 84% identity to bovine EFIA#1. The genomic cloning data indicate that processed pseudogenes are partially responsible for the complexity of the EFIA genomic Southern blots. The phenomenon of 'repeat induced point mutation' (ripping) at bovine psi EFIA#1 gene CpG dinucleotides occurs at a 6.5-fold higher frequency than expected from random mutagenesis. Therefore, ripping is likely to be the mechanism by which the bovine EFIA#1 pseudogene's ectopic recombination potential was inactivated.
已对CCAAT转录复合体组分增强子因子I亚基A(EFIA)的基因组拷贝多样性进行了检测。当用大鼠EFIA cDNA与哺乳动物基因组Southern印迹杂交时,在所检测的所有物种中均发现了由众多相关序列组成的复杂图谱,其中牛的图谱最为简单。通过用1489 bp的大鼠EFIA cDNA进行筛选,从原代肺内皮细胞cDNA文库中分离出了牛的EFIA#1 cDNA。推导的牛EFIA#1氨基酸(aa)序列与大鼠EFIA的同源性为98%,与人类EFIA/DbpB/YB-1家族成员DNA结合蛋白B(DbpB)的同源性为100%。此外,通过用大鼠EFIA cDNA探针筛选基因组文库,获得了牛的一个加工过的EFIA假基因,命名为牛psi EFIA#1。牛psi EFIA#1基因有一个开放阅读框(ORF),如果表达,将编码一个140个氨基酸的序列,其中第31至140位氨基酸与牛EFIA#1的同源性为84%。基因组克隆数据表明,加工过的假基因部分导致了EFIA基因组Southern印迹的复杂性。牛psi EFIA#1基因CpG二核苷酸处的“重复诱导点突变”(ripping)现象发生频率比随机诱变预期的高6.5倍。因此,ripping可能是使牛EFIA#1假基因的异位重组潜能失活的机制。