Albig W, Bramlage B, Gruber K, Klobeck H G, Kunz J, Doenecke D
Institut für Biochemie und Molekulare Zellbiologie, Georg-August-Universität Göttingen, Germany.
Genomics. 1995 Nov 20;30(2):264-72. doi: 10.1006/geno.1995.9878.
H3.3 is a replacement histone subtype that is encoded by two replication-independent genes termed H3.3A and H3.3B, respectively. We have isolated a fullsize H3.3 cDNA clone from an oligo(dT)-primed human testicular cDNA library. Subsequently, the corresponding gene was isolated from a human cosmid library and was identified as the H3.3B gene. It was the only histone gene on this 42-kb cosmid clone. The gene structure shows characteristic features of an H3.3 gene. First, it contains an intron of about 0.5 kb in the 5' untranslated region and two smaller introns within the coding gene portion. Second, no histone gene-specific dyad symmetry element was found in the 3' untranslated region, but three putative polyadenylation signals were detected downstream of the gene. The corresponding transcripts were detected by Northern blot analysis using poly(A)+ RNA from testis and from the HEK293 tumor cell line. The newly discovered human H3.3B gene (HGMW-approved symbol H3F3B) was mapped by fluorescence in situ hybridization to the telomeric region of chromosome 17 (17q25). This localization of the H3.3B gene and its solitary arrangement contrast with the majority of the replication-dependent histone genes, which form a large cluster on chromosome 6 and a second cluster on chromosome 1.
H3.3是一种替换组蛋白亚型,分别由两个不依赖复制的基因H3.3A和H3.3B编码。我们从一个oligo(dT)引物介导的人睾丸cDNA文库中分离出一个全长H3.3 cDNA克隆。随后,从人黏粒文库中分离出相应基因,并鉴定为H3.3B基因。它是这个42kb黏粒克隆上唯一的组蛋白基因。该基因结构显示出H3.3基因的特征。首先,它在5'非翻译区含有一个约0.5kb的内含子,在编码基因部分内有两个较小的内含子。其次,在3'非翻译区未发现组蛋白基因特异性的二元对称元件,但在基因下游检测到三个推定的聚腺苷酸化信号。使用来自睾丸和HEK293肿瘤细胞系的poly(A)+RNA通过Northern印迹分析检测到相应的转录本。通过荧光原位杂交将新发现的人H3.3B基因(HGMW批准的符号H3F3B)定位到染色体17的端粒区域(17q25)。H3.3B基因的这种定位及其单独排列与大多数依赖复制的组蛋白基因形成对比,后者在染色体6上形成一个大簇,在染色体1上形成第二个簇。