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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development
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Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development

机译：男性生殖细胞发育过程中蛋白激酶A催化亚基非肉豆蔻酰化变体的表达

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摘要

The catalytic subunits of protein kinase A are transcribed in all mouse tissues from two distinct genes that code for the C_α and C_β isoforms. Alternative promoters exist for the C_β gene that are used in a tissue-specific fashion and give rise to variants that differ in their amino-terminal sequences. We have characterized an alternative promoter that is present in the first intron of the C_α gene and is transcriptionally active in male germ cells. Transcription from this promoter is coincident with the appear- ance of pachytene spermatocytes and leads to a C_α protein (C_α2) that contains a distinctive 7 amino acid amino-terminus differing from the 14 amino acid amino-terminus of C_α1. The C_α2 protein does not contain the myristylation signal present on C_α1 and migrates at a lower molecular weight on SDS/PAGE gels. By Western blotting, we estimate that most or all of the C_α protein present in mature sperm is C_α2. The amino-terminal sequence of Ca2 is similar to that of ovine sperm C as previously reported [San Agustin. J. T., Leszyk, J. D.. Nuwaysir, L. M. & Witman. G. B. (1998) i. Biol. Chem. 273. 24874-24883], and we show by cDNA cloning that human sperm also express a highly related C_α2 homolog. The Co2 subunit forms holoenzymes with either Rll_α or RIa, and both activate at the same concentration of cyclic nucleotide. Because protein kinase A is thought to play a pivotal role in sperm motility and capacitation, the distinctive biochem- ical properties of the unmyristylated C_α2 may be essential for fertility in the male.
机译：蛋白激酶A的催化亚基从编码C_α和C_β同工型的两个不同基因转录到所有小鼠组织中。存在以组织特异性方式使用的C_β基因的替代启动子，并产生其氨基末端序列不同的变体。我们已经表征了存在于C_α基因的第一内含子中并且在雄性生殖细胞中具有转录活性的替代启动子。该启动子的转录与粗线精子细胞的出现相吻合，并导致C_α蛋白（C_α2）含有与C_α1的14个氨基酸不同的7个氨基酸的氨基末端。 C_α2蛋白不含C_α1上存在的肉豆蔻酰化信号，并以较低的分子量在SDS / PAGE凝胶上迁移。通过蛋白质印迹，我们估计成熟精子中存在的大部分或全部C_α蛋白是C_α2。 Ca2的氨基末端序列类似于先前报道的绵羊精子C的序列[San Agustin。 J. T.，Leszyk，J。D. Nuwaysir，L。M.和Witman。湾（1998）i。生物学化学273. 24874-24883]，并且我们通过cDNA克隆证明人精子也表达高度相关的C_α2同源物。 Co2亚基与Rll_α或RIa形成全酶，并且都以相同浓度的环状核苷酸激活。由于人们认为蛋白激酶A在精子运动和获能中起着关键作用，因此，非肉豆蔻酰化的C_α2的独特生化特性可能对男性的生育能力至关重要。

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