Khanna A, Campbell R D
Department of Biochemistry, University of Oxford, U.K.
Biochem J. 1996 Oct 1;319 ( Pt 1)(Pt 1):81-9. doi: 10.1042/bj3190081.
G13 is a single-copy gene lying approx. 75 kb centromeric of the complement gene cluster in the class III region of the human MHC. The gene spans approx. 17 kb of DNA and has been shown to encode mRNA of approx. 2.7 kb that is present in cell lines representing lymphoid and non-lymphoid tissues, indicating that it is ubiquitously expressed. The complete nucleotide sequence of the 2.7 kb mRNA has been derived from cDNA and genomic clones. The longest open reading frame obtained for G13 codes for a 703 amino acid protein of approx. 77 kDa in molecular mass. Comparison of the putative G13 amino acid sequence with the protein databases revealed significant similarities with DNA-binding proteins of the leucine zipper class, including a human cAMP response element binding protein. G13 contains a bZIP motif, a region rich in basic amino acids adjacent to a coiled-coil leucine zipper domain, common to this class of proteins that is known to be involved in dimerization and DNA binding. Antibodies raised against a fragment encoding the C-terminal half of the putative G13 protein recognized a major polypeptide of approx. 86 kDa and a minor polypeptide of approx. 78 kDa on immunoblotting of U937 cell extracts; this has been confirmed by immunoprecipitation experiments. Even though it contained at least one potential bipartite nuclear localization signal, the G13 protein was present both in the cytoplasm and the nucleus of the fibroblast cells. Thus G13 might be a novel DNA-binding protein that is perhaps translocated to the nucleus in a regulated manner.
G13是一个单拷贝基因,位于人类MHC III类区域补体基因簇着丝粒侧约75 kb处。该基因跨越约17 kb的DNA,已显示其编码约2.7 kb的mRNA,在代表淋巴组织和非淋巴组织的细胞系中均有表达,表明它在全身广泛表达。2.7 kb mRNA的完整核苷酸序列已从cDNA和基因组克隆中获得。从G13获得的最长开放阅读框编码一个约703个氨基酸的蛋白质,分子量约为77 kDa。将推测的G13氨基酸序列与蛋白质数据库进行比较,发现它与亮氨酸拉链类的DNA结合蛋白有显著相似性,包括一种人类cAMP反应元件结合蛋白。G13包含一个bZIP基序,即一个富含碱性氨基酸的区域,与这类蛋白质中常见的卷曲螺旋亮氨酸拉链结构域相邻,已知该结构域参与二聚化和DNA结合。针对编码推测的G13蛋白C端一半的片段产生的抗体,在对U937细胞提取物进行免疫印迹时识别出一条约86 kDa的主要多肽和一条约78 kDa的次要多肽;免疫沉淀实验已证实了这一点。尽管G13蛋白含有至少一个潜在的双分型核定位信号,但它在成纤维细胞的细胞质和细胞核中均有存在。因此,G13可能是一种新型的DNA结合蛋白,可能以一种受调控的方式转运到细胞核中。