Skeletor, a novel chromosomal protein that redistributes during mitosis provides evidence for the formation of a spindle matrix.
A spindle matrix has been proposed to help organize and stabilize the microtubule spindle during mitosis, though molecular evidence corroborating its existence has been elusive. In Drosophila, we have cloned and characterized a novel nuclear protein, skeletor, that we propose is part of a macromolecular complex forming such a spindle matrix. Skeletor antibody staining shows that skeletor is associated with the chromosomes at interphase, but redistributes into a true fusiform spindle structure at prophase, which precedes microtubule spindle formation. During metaphase, the spindle, defined by skeletor antibody labeling, and the microtubule spindles are coaligned. We find that the skeletor-defined spindle maintains its fusiform spindle structure from end to end across the metaphase plate during anaphase when the chromosomes segregate. Consequently, the properties of the skeletor-defined spindle make it an ideal substrate for providing structural support stabilizing microtubules and counterbalancing force production. Furthermore, skeletor metaphase spindles persist in the absence of microtubule spindles, strongly implying that the existence of the skeletor-defined spindle does not require polymerized microtubules. Thus, the identification and characterization of skeletor represents the first direct molecular evidence for the existence of a complete spindle matrix that forms within the nucleus before microtubule spindle formation.
Walker
D L
DL
Department of Zoology and Genetics, Iowa State University, Ames, Iowa 50011, USA.
Wang
D
D
Jin
Y
Y
Rath
U
U
Wang
Y
Y
Johansen
J
J
Johansen
K M
KM
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001483:Base Sequence; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000737:chemistry / Q000166:cytology* / Q000196:embryology / Q000235:genetics*; D007150:Immunohistochemistry; D008938:Mitosis; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D034521:Nuclear Matrix-Associated Proteins; D016366:Open Reading Frames / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D016415:Sequence Alignment; D008941:Spindle Apparatus / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
D000906:Antibodies; D002868:Chromosomal Proteins, Non-Histone; D029721:Drosophila Proteins; D034521:Nuclear Matrix-Associated Proteins; D012333:RNA, Messenger; C419556:skeletor protein, Drosophila; D015739:Nocodazole