Nek2B stimulates zygotic centrosome assembly in Xenopus laevis in a kinase-independent manner.
Pronuclear migration and formation of the first mitotic spindle depend upon assembly of a functional zygotic centrosome. For most animals, this involves both paternal and maternal contributions as sperm basal bodies are converted into centrosomes competent for microtubule nucleation through recruitment of egg proteins. Nek2B is a vertebrate NIMA-related protein kinase required for centrosome assembly, as its depletion from egg extracts delays microtubule aster formation from sperm basal bodies. Using Xenopus as a model system, we now show that protein expression of Nek2B begins during mid-oogenesis and increases further upon oocyte maturation. This is regulated, at least in part, at the level of protein translation. Nek2B protein is weakly phosphorylated in mitotic egg extracts but its recruitment to the sperm basal body, which occurs independently of its kinase activity, stimulates its phosphorylation, possibly through sequestration from a phosphatase present in mitotic egg cytoplasm. Importantly, although Nek2B is not required to organize acentrosomal microtubule asters, we show that addition of either active or kinase-dead recombinant Nek2B can restore centrosome assembly in a dose-dependent manner to a depleted extract. These results support a model in which maternal Nek2B acts to promote assembly of a functional zygotic centrosome in a kinase-independent manner.
Twomey
Ciara
C
Department of Biochemistry, University of Leicester, University Road, Leicester LE1 7RH, UK.
Wattam
Samantha L
SL
Pillai
Meenu R
MR
Rapley
Joe
J
Baxter
Joanne E
JE
Fry
Andrew M
AM
D000818:Animals; D018385:Centrosome / Q000201:enzymology* / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D005260:Female; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008870:Microtubules / Q000201:enzymology; D009865:Oocytes / Q000201:enzymology; D009866:Oogenesis / Q000502:physiology; D010063:Ovum / Q000201:enzymology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D013094:Spermatozoa / Q000201:enzymology; D029867:Xenopus Proteins; D014982:Xenopus laevis; D015053:Zygote / Q000201:enzymology* / Q000378:metabolism
D007527:Isoenzymes; D029867:Xenopus Proteins; C490316:NEK2 protein, Xenopus; C490315:Nek2B protein, Xenopus; D017346:Protein Serine-Threonine Kinases