Harduin-Lepers A, Shaper N L, Mahoney J A, Shaper J H
Cell Structure and Function Laboratory, Johns Hopkins University School of Medicine, Baltimore, MD 21287.
Glycobiology. 1992 Aug;2(4):361-8. doi: 10.1093/glycob/2.4.361.
We have previously shown that the expression of the gene encoding murine beta 1,4-galactosyltransferase (beta 1,4-GT, UDP-galactose:N-acetyl-D-glucosaminyl-glycopeptide 4-beta-D galactosyltransferase, EC 2.4.1.38) is fundamentally different between somatic and male germ cells (Shaper et al., 1990b). In somatic cells, two transcripts of 3.9 kb and 4.1 kb are produced. In contrast, in spermatogonia only the 4.1 kb transcript is expressed. Maturation of spermatogonia to pachytene spermatocytes is accompanied by reduced expression of the 4.1 kb transcript to barely detectable levels. Continued differentiation to haploid round spermatids is coincident with renewed expression in which the 4.1 kb transcript is replaced by two truncated transcripts of 2.9 and 3.1 kb. In this study, we report the characterization of a full-length beta 1,4-GT cDNA clone from a murine round spermatid library that corresponds to the 2.9 kb transcript. This transcript encodes the same open reading frame as the 4.1 kb transcript, but utilizes alternative poly(A) signals embedded within the long 3'-untranslated region of the somatic transcript. Based on sequence analysis, together with primer extension and S1 nuclease protection experiments, both the 2.9 and the 3.1 kb round spermatid beta 1,4-GT transcripts are distinguished by the presence of an additional 5'-untranslated sequence of approximately 560 bp that is absent in premeiotic germ cells and somatic cells.(ABSTRACT TRUNCATED AT 250 WORDS)
我们之前已经表明,编码小鼠β1,4-半乳糖基转移酶(β1,4-GT,UDP-半乳糖:N-乙酰-D-葡糖胺基糖肽4-β-D-半乳糖基转移酶,EC 2.4.1.38)的基因在体细胞和雄性生殖细胞中的表达存在根本差异(Shaper等人,1990b)。在体细胞中,产生3.9 kb和4.1 kb的两种转录本。相比之下,在精原细胞中仅表达4.1 kb的转录本。精原细胞向粗线期精母细胞的成熟伴随着4.1 kb转录本的表达降低至几乎检测不到的水平。持续分化为单倍体圆形精子细胞与重新表达同时发生,其中4.1 kb转录本被2.9 kb和3.1 kb的两种截短转录本所取代。在本研究中,我们报告了从小鼠圆形精子细胞文库中获得的与2.9 kb转录本相对应全长β1,4-GT cDNA克隆的特征。该转录本与4.1 kb转录本编码相同的开放阅读框,但利用了体细胞转录本长3'-非翻译区内的替代聚腺苷酸化信号。基于序列分析,以及引物延伸和S1核酸酶保护实验,2.9 kb和3.1 kb圆形精子细胞β1,4-GT转录本的特征均在于存在约560 bp的额外5'-非翻译序列,该序列在减数分裂前的生殖细胞和体细胞中不存在。(摘要截断于250字)