Phosphorylation of the major Drosophila lamin in vivo: site identification during both M-phase (meiosis) and interphase by electrospray ionization tandem mass spectrometry.
Phosphorylation can have profound effects on the properties of nuclear lamins. For instance, phosphorylation of specific sites on mammalian lamins drastically alters their propensity to polymerize. Relatively little is known about the effects of phosphorylation during interphase and about phosphorylation of invertebrate nuclear lamins. Here, using electrospray ionization tandem mass spectrometry, we determined the phosphorylation sites of both interphase and M-phase isoforms of nuclear lamin Dm from Drosophila melanogaster. Interphase lamins are phosphorylated at three sites: two of these sites (Ser25 and a site located between residues 430 and 438) flank the alpha-helical rod domain, whereas the third site (Ser595) is located close to the C-terminus. The M-phase lamin isoform is phosphorylated predominantly at Ser45, a residue contained within a sequence matching the consensus site for phosphorylation by cdc2 kinase. Our study confirms the important role in vivo for cdc2 kinase in M-phase disassembly of nuclear lamins and provides the basis for understanding Drosophila lamin phosphorylation during interphase.
Schneider
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U
Department of Biochemistry, Biozentrum of the University of Basel, Switzerland.
Mini
T
T
Jenö
P
P
Fisher
P A
PA
Stuurman
N
N
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D004331:Drosophila melanogaster / Q000737:chemistry*; D006801:Humans; D007399:Interphase; D034882:Lamins; D013058:Mass Spectrometry / Q000379:methods; D008540:Meiosis; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D010449:Peptide Mapping / Q000379:methods; D010748:Phosphopeptides / Q000032:analysis; D010766:Phosphorylation
D034882:Lamins; D009687:Nuclear Proteins; D010748:Phosphopeptides