Mori C, Welch J E, Fulcher K D, O'Brien D A, Eddy E M
Gamete Biology Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709.
Biol Reprod. 1993 Aug;49(2):191-203. doi: 10.1095/biolreprod49.2.191.
We have identified cDNAs representing three hexokinase mRNAs (Hk1-sa, Hk1-sb, Hk1-sc) by screening mouse spermatogenic cell cDNA libraries with a mouse hepatoma cell line hexokinase (Hk1) cDNA [Arora KK, Fanciulli M, Pederson PL. J Biol Chem 1990; 265:6481-6488]. Although all three cDNAs show 99% identity to the somatic Hk1 cDNA sequence throughout most of their coding region, they differ from this sequence at the 5' end. They contain a common spermatogenic cell-specific sequence and a sequence unique to each cDNA immediately 5' to the common domain. However, they lack the porin-binding domain (PBD) present in this region of Hk1, used for binding to a pore-forming protein in the outer mitochondrial membrane. These observations appear to support a model proposed by others for hexokinase gene evolution in mammals. In addition, we found that Hk1-sb has an internal sequence that is not present in Hk1, Hk1-sa, or Hk1-sc. Moreover, Hk1-sa and Hk1-sb transcripts are developmentally expressed in mouse spermatogenic cells. Hk1-sa mRNA is first expressed during meiosis and continues to be present in postmeiotic germ cells, while the more abundant Hk1-sb mRNA is detected only in postmeiotic germ cells. These and other findings suggest that enzymes encoded by Hk1-sa, Hk1-sb, and Hk1-sc are present only in spermatogenic cells.
我们通过用小鼠肝癌细胞系己糖激酶(Hk1)cDNA [Arora KK, Fanciulli M, Pederson PL. J Biol Chem 1990; 265:6481 - 6488] 筛选小鼠生精细胞cDNA文库,鉴定出了代表三种己糖激酶mRNA(Hk1 - sa、Hk1 - sb、Hk1 - sc)的cDNA。尽管所有这三种cDNA在其大部分编码区域与体细胞Hk1 cDNA序列显示出99%的同一性,但它们在5'端与该序列不同。它们包含一个共同的生精细胞特异性序列以及在共同结构域紧邻5'端的每个cDNA特有的序列。然而,它们缺乏Hk1该区域中存在的用于与线粒体外膜中一种成孔蛋白结合的孔蛋白结合结构域(PBD)。这些观察结果似乎支持了其他人提出的关于哺乳动物己糖激酶基因进化的模型。此外,我们发现Hk1 - sb有一个Hk1、Hk1 - sa或Hk1 - sc中不存在的内部序列。而且,Hk1 - sa和Hk1 - sb转录本在小鼠生精细胞中呈发育性表达。Hk1 - sa mRNA在减数分裂期间首次表达,并在减数分裂后生殖细胞中持续存在,而更丰富的Hk1 - sb mRNA仅在减数分裂后生殖细胞中被检测到。这些以及其他发现表明,由Hk1 - sa、Hk1 - sb和Hk1 - sc编码的酶仅存在于生精细胞中。