Molecular characterisation of cAMP-dependent protein kinase (PK-A) catalytic subunit isoforms in the male tick, Amblyomma hebraeum

被引:9
作者
Tabish, Mohammad
Clegg, Roger A.
Turner, Philip C.
Jonczy, Jan
Rees, Huw H.
Fisher, Michael J.
机构
[1] Univ Liverpool, Cellular Regulat & Signalling Grp, Sch Biol Sci, Liverpool L69 7ZB, Merseyside, England
[2] Hannah Res Inst, Ayr KA6 5HL, Scotland
[3] Ctr Rech St Anim SA, Farm Anim Parasitiides, CH-1566 St Aubin, Switzerland
基金
英国生物技术与生命科学研究理事会;
关键词
tick; isoform; transcript; sperm; protein kinase; Amblyomma hebraeum;
D O I
10.1016/j.molbiopara.2006.09.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The cAMP-dependent protein kinase (protein kinase A, PK-A) plays a central role in the regulation of diverse aspects of cellular activity. Specifically, PK-A appears to play a key controlling role in the maturation of spermatids. Using a PCR-based approach, with degenerate primers from the highly conserved regions of the PK-A catalytic (C) subunit in combination with 5' and 3' RACE, we have cloned three cDNAs for the PK-A C-subunit of the male tick, Amblyomma hebraeum. The three cDNAs have open reading frames of 1059, 1275 and 1404 bp which encode proteins of 40.6, 48.2 and 52.5 kDa, respectively. These transcripts appear to arise from 5' alternative splicing of RNA derived from a single gene for the PK-A C-subunit. One isoform (AH-PK-A C1), in common with PK-A C-subunits from a range of species, contains a consensus sequence for N-myristoylation. RT-PCR and Western blot experiments suggest that the three splice variants are expressed ubiquitously; however, expression of the myristoylatable AH-PK-A C1 isoform is predominant in all investigated tissues (accessory gland, midgut, Malpighian tubules, salivary gland, testis and immature spermatids). There is no evidence for a sperm-specific PK-A C-subunit (Cs) in tick sperm; however, tyrosine protein phosphorylation, previously shown to be modulated by PK-A activity during mammalian sperm maturation, was observed in tick sperm. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:330 / 339
页数:10
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