Giorda R, Ohmachi T, Ennis H L
Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110.
J Mol Biol. 1989 Jan 5;205(1):63-9. doi: 10.1016/0022-2836(89)90364-1.
mRNA specific to cDNA clone pLK109 is present in Dictyostelium discoideum spores, increases about two- to threefold at 0.5 to 1 h during spore germination, and then rapidly decreases. The mRNA is not detectable in vegetative cells or in early multicellular development on filters, but is present late during development, approximately at the time of sporulation. 109 mRNA in spores is 700 nucleotides in length but this is processed during germination by shortening of the poly(A) tail to about 600 nucleotides at 1 to 1.5 hours. pLK109 is a member of a multigene family containing three separate genes, and we have isolated and sequenced all of them. All three sequences code for deduced proteins of 127 amino acid residues, with only a few amino acid differences among them. Gene 1 represents the "transcribed" gene, since all 33 cDNAs we isolated are identical with the cDNA pLK109 and the coding region of this gene. Other open reading frames are in close proximity to each of the 109 sequences. About 200 base-pairs 3' to the gene 1 109 sequence is an open reading frame in the opposite orientation. Gene 2 fragment contains a sequence that codes for a protein similar to trypanosome alpha-tubulin 728 base-pairs 5' to the 109 sequence. Gene 3 fragment possesses two additional putative coding regions, one 5' and another 3' to the 109 gene. There is a remarkable similarity between the 5' upstream regions of all three genes. Each possesses a normal Dictyostelium TATA box and the usual T stretch. In addition, there are many other portions of about 400 to 500 base-pairs of the 5' regions that are either identical for long stretches or very similar.
特定于cDNA克隆pLK109的mRNA存在于盘基网柄菌的孢子中,在孢子萌发的0.5至1小时内增加约两到三倍,然后迅速下降。在营养细胞或滤膜上的早期多细胞发育阶段无法检测到该mRNA,但在发育后期,大约在孢子形成时存在。孢子中的109 mRNA长度为700个核苷酸,但在萌发过程中通过将多聚腺苷酸尾巴在1至1.5小时缩短至约600个核苷酸进行加工。pLK109是一个包含三个独立基因的多基因家族的成员,我们已经分离并对它们全部进行了测序。所有三个序列编码推导的127个氨基酸残基的蛋白质,它们之间只有少数氨基酸差异。基因1代表“转录”基因,因为我们分离的所有33个cDNA与cDNA pLK109以及该基因的编码区相同。其他开放阅读框与每个109序列紧密相邻。在基因1的109序列3'端约200个碱基对处是一个反向的开放阅读框。基因2片段包含一个编码与锥虫α-微管蛋白相似的蛋白质的序列,位于109序列5'端728个碱基对处。基因3片段有两个额外的推定编码区,一个在109基因的5'端,另一个在3'端。所有三个基因的5'上游区域之间存在显著相似性。每个都有一个正常的盘基网柄菌TATA盒和通常的T序列。此外,5'区域约400至500个碱基对的许多其他部分要么有很长一段相同,要么非常相似。