PROTEIN AND NUCLEAR-CHANGES IN PIG EGGS AT FERTILIZATION

被引:43
作者
DING, JC
CLARKE, N
NAGAI, T
MOOR, RM
机构
[1] AFRC, INST ANIM PHYSIOL & GENET RES, DEPT MOLEC EMBRYOL, CAMBRIDGE CB2 4AT, ENGLAND
[2] NATL INST ANIM IND, NORINDANCHI, JAPAN
关键词
PHOSPHORYLATION; PROTEIN SYNTHESIS; PRONUCLEI;
D O I
10.1002/mrd.1080310410
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 nuclear restructuring that occurs between insemination and full pronuclear formation in pig eggs is accompanied by posttranslational changes to specific egg proteins. Sperm penetration begins in vitro at 3 hr postinsemination (hpi). By 5 hr, decondensing sperm heads and anaphase II plates are observed in 50% of eggs, and, by 8 hpi, both male and female pronuclei have formed. Three consistent changes to the pattern of newly synthesised proteins are triggered in this period; they affect the 46K, 25K, and 22K polypeptides. Changes are also triggered in the 180-200K polypeptides and in the 14K polypeptides, but these are highly variable. The same changes in the prefertilization pattern were observed when prelabelled eggs were used and new protein synthesis was suppressed. The first and most abrupt change involves the apparent catabolic elimination of a group of 46K unphosphorylated polypeptides (pI 7.3-6.4), whose synthesis was greatest before germinal vesicle breakdown but declined slowly in the final phase of maturation, then declined precipitously after activation. Ageing (beyond maturation) also leads to the disappearance of these polypeptides. The progressive disappearance of a set of 25K polypeptides and the concomitant appearance of a dominant 22K polypeptide is the most characteristic fertilization-induced modification to porcine egg proteins. These modifications begin within 1 hr of sperm penetration or activation, are specific to the pig, and involve heavily phosphorylated polypeptides (25K, pI 6.7-6.0) whose synthesis is begun in the early metaphase I stage. Dual ([S-35] and [P-32]) labelling, protein blocking experiments, and use of alkaline phosphatase suggest that dephosphorylation selectively affects these 25K polypeptides and is mainly or wholly responsible for converting them (completely within 6 hr) to a single, new (22K, pI 7.6) species that is positively charged. The 25K/22K polypeptide modification has a close temporal relationship with the formation of the male and female pronuclei.
引用
收藏
页码:287 / 296
页数:10
相关论文
共 25 条
[1]   FILM DETECTION METHOD FOR TRITIUM-LABELED PROTEINS AND NUCLEIC-ACIDS IN POLYACRYLAMIDE GELS [J].
BONNER, WM ;
LASKEY, RA .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1974, 46 (01) :83-88
[2]   CAPACITATION OF RABBIT SPERMATOZOA INVITRO [J].
BRACKETT, BG ;
OLIPHANT, G .
BIOLOGY OF REPRODUCTION, 1975, 12 (02) :260-274
[3]  
CHENG WTK, 1985, THESIS AFRC I ANIMAL
[4]  
DING J, 1988, J REPR FERT ABSTR SE, V2, P23
[5]   IONOMYCIN-REGULATED PHOSPHORYLATION OF THE MYELOID CALCIUM-BINDING PROTEIN P14 [J].
EDGEWORTH, J ;
FREEMONT, P ;
HOGG, N .
NATURE, 1989, 342 (6246) :189-192
[6]   STAGE-SPECIFIC CHANGES IN PROTEIN-PHOSPHORYLATION ACCOMPANYING MEIOTIC MATURATION OF MOUSE OOCYTES AND FERTILIZATION OF MOUSE EGGS [J].
ENDO, Y ;
KOPF, GS ;
SCHULTZ, RM .
JOURNAL OF EXPERIMENTAL ZOOLOGY, 1986, 239 (03) :401-409
[7]  
FULKA J, 1986, J REPROD FERTIL, V77, P281, DOI 10.1530/jrf.0.0770281
[8]  
GARRISON JC, 1982, J BIOL CHEM, V257, P3135
[9]   DEPHOSPHORYLATION AND ACTIVATION OF XENOPUS-P34CDC2 PROTEIN-KINASE DURING THE CELL-CYCLE [J].
GAUTIER, J ;
MATSUKAWA, T ;
NURSE, P ;
MALLER, J .
NATURE, 1989, 339 (6226) :626-629
[10]  
HOWLETT SK, 1985, J EMBRYOL EXP MORPH, V87, P175