Two proteins that cycle asynchronously between centrosomes and nuclear structures: Drosophila CP60 and CP190.
Both the nucleus and the centrosome are complex, dynamic structures whose architectures undergo cell cycle-specific rearrangements. CP190 and CP60 are two Drosophila proteins of unknown function that shuttle between centrosomes and nuclei in a cell cycle-dependent manner. These two proteins are associated in vitro, and localize to centrosomes in a microtubule independent manner. We injected fluorescently labeled, bacterially expressed CP190 and CP60 into living Drosophila embryos and followed their behavior during the rapid syncytial blastoderm divisions (nuclear cycles 10-13). Using quantitative 3-D wide-field fluorescence microscopy, we show that CP190 and CP60 cycle between nuclei and centrosomes asynchronously with the accumulation of CP190 leading that of CP60 both at centrosomes and in nuclei. During interphase, CP190 is found in nuclei. Immediately following nuclear envelope breakdown, CP190 localizes to centrosomes where it remains until telophase, thereafter accumulating in reforming nuclei. Unlike CP190, CP60 accumulates at centrosomes primarily during anaphase, where it remains into early interphase. During nuclear cycles 10 and 11, CP60 accumulates in nuclei simultaneous with nuclear envelope breakdown, suggesting that CP60 binds to an unknown nuclear structure that persists into mitosis. During nuclear cycles 12 and 13, CP60 accumulates gradually in nuclei during interphase, reaching peak levels just before nuclear envelope breakdown. Once in the nucleus, both CP190 and CP60 appear to form fibrous intranuclear networks that remain coherent even after nuclear envelope breakdown. The CP190 and CP60 networks do not co-localize extensively with each other or with DNA. This work provides direct evidence, in living cells, of a coherent protein network that may represent a nuclear skeleton.
Oegema
K
K
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448, USA.
Marshall
W F
WF
Sedat
J W
JW
Alberts
B M
BM
D000818:Animals; D018797:Cell Cycle Proteins; D018385:Centrosome / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000032:analysis; D004330:Drosophila / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D015530:Nuclear Matrix / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016335:Zinc Fingers / Q000502:physiology
D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; C098388:Map60 protein, Drosophila; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA