Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family
Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family
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DOI：
10.4238/2015.october.18.56
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发表时间：
2015-01-01
影响因子：
0.4
通讯作者：
Xiao, H.
中科院分区：
文献类型：
--
作者：
Wang, P.;Huo, H. L.;Xiao, H.
睾丸特异性丝氨酸激酶（tsks）是一个在睾丸中高度表达的丝氨酸/苏氨酸激酶家族，负责调节许多与精子发生相关的蛋白质活性。该家族突变与人类和小鼠少精子症和无精子症呈正相关。本文对来自版纳迷你猪自交系（BMI）的5个TSSK家族成员进行了克隆、测序和鉴定。BMI TSSKs的全长编码序列在807 ~ 1095 bp之间，编码268 ~ 364个氨基酸，分子量在30.11 ~ 41.34 kDa之间。在与其他物种的TSSK4基因进行比较后，发现BMI TSSK4包含三个可变剪接变体，inform1、inform 3和inform 4。BMI TSSK1和TSSK2共定位于Sus scrofa染色体（SSC） 14上，由单个外显子组成；TSSK3、TSSK4和TSSK6位于SSC6、SSC7和SSC2上，分别由2个、4个和1个外显子组成。多个蛋白序列比对和系统发育分析表明，跨越S_TKc结构域的区域在猪和其他动物之间更为保守：TSSK1和TSSK2、TSSK3和TSSK6在物种间的同源性最大，TSSK4蛋白与其他成员蛋白明显不同。多组织RT-PCR结果显示，BMI TSSK1、TSSK3和TSSK4仅在睾丸和精囊中表达，TSSK2仅局限于睾丸，而TSSK6在成人组织中广泛表达，但在睾丸中表达量最高。
Testis-specific serine kinases (TSSKs) are a family of serine/threonine kinases highly expressed in the testes that are responsible for regulating many spermatogenesis-related protein activities. Mutations in this family have a positive relationship with oligospermia and azoospermia in human and mouse. Here, five members of the TSSK family from a Banna mini-pig inbred line (BMI) were cloned, sequenced, and characterized. The full-length coding sequences of BMI TSSKs varied from 807 (TSSK3) to 1095 bp (TSSK1) and encoded 268 to 364 amino acids with molecular weights in the range 30.11 to 41.34 kDa. Following comparison with TSSK4 genes in other species, BMI TSSK4 was found to contain three alternatively spliced variants, inform1, inform 3, and inform 4. BMI TSSK1 and TSSK2 are co-localized on the Sus scrofa chromosome (SSC) 14, and consist of a single exon; TSSK3, TSSK4, and TSSK6 are on SSC6, SSC7, and SSC2, and consist of two, four, and one exon, respectively. Multiple protein sequence alignment and phylogenetic analysis showed that the regions spanning the S_TKc domains were more conserved between pig and other animals: with TSSK1 and TSSK2 and TSSK3 and TSSK6 displaying the greatest degree of homology across species, and the TSSK4 protein clearly distinct from other members. Multi-tissue RT-PCR showed BMI TSSK1, TSSK3, and TSSK4 were only expressed in the testes and seminal vesicle, TSSK2 was confined to testes only, while TSSK6 was expressed widely in adult tissues but was highest in the testes.