TY - JOUR
T1 - Cell cycle-dependent phosphorylation of centrosomes
T2 - Localization of phosphopeptide specific antibodies to the centrosome
AU - Vandré, Dale D.
AU - Feng, Yang
AU - Ding, Min
N1 - Funding Information:
Supported by Grant No. DK 33225 from the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health (NIH); Grant No. M01 RR 00400 from the NIH General Clinical Research Center at the University of Minnesota; the Minnesota Medical Foundation; and the Diabetes Trust Fund, Inc, Birmingham, Alabama, Buris R. Boshell, MD, founder. W. H. was supported by an American Diabetes Association Career Development Award and by a Clinical Associate Physician Award of the University of Washington General Clinical Research Center (NIH Grant No. RR 00037).
PY - 2000/6/1
Y1 - 2000/6/1
N2 - The microtubule nucleation capacity of the centrosome increases dramatically as cells progress from interphase into mitosis. The increase in nucleation capacity of the centrosome correlates with the cell cycle- dependent localization of the mitotic protein monoclonal-2 (MPM-2) phosphoepitope-specifc antibody to the mitotic centrosome. Therefore, the phosphorylation state of centrosomal components may regulate the microtubule nucleation capacity of this organelle during mitosis. Neither the identity of the MPM-2 kinase(s) nor all of the MPM-2-reactive phosphoproteins associated with the centrosome have been fully elucidated. Only recently have the characteristics of the MPM-2 epitope site been defined, and we used this information to prepare polyclonal antibodies against synthetic phosphopeptides containing potential MPM-2 epitopes derived from the sequences of two MPM-2-reactive proteins, topoisomerase II, and microtubule associated protein 1B (MAP1B). We demonstrate that these phosphopeptide- specific antibodies also localize to the centrosome in a cell cycle-dependent fashion. Thus, polyclonal antibodies have been generated against defined phosphopeptides that reiterate many of the immunofluorescence staining properties exhibited by the MPM-2 antibody. These new phosphopeptide-specific antibodies will provide additional probes to examine the phosphorylation of centrosomal components and the functional consequences of their phosphorylation during mitosis. (C) 2000 Wiley-Liss, Inc.
AB - The microtubule nucleation capacity of the centrosome increases dramatically as cells progress from interphase into mitosis. The increase in nucleation capacity of the centrosome correlates with the cell cycle- dependent localization of the mitotic protein monoclonal-2 (MPM-2) phosphoepitope-specifc antibody to the mitotic centrosome. Therefore, the phosphorylation state of centrosomal components may regulate the microtubule nucleation capacity of this organelle during mitosis. Neither the identity of the MPM-2 kinase(s) nor all of the MPM-2-reactive phosphoproteins associated with the centrosome have been fully elucidated. Only recently have the characteristics of the MPM-2 epitope site been defined, and we used this information to prepare polyclonal antibodies against synthetic phosphopeptides containing potential MPM-2 epitopes derived from the sequences of two MPM-2-reactive proteins, topoisomerase II, and microtubule associated protein 1B (MAP1B). We demonstrate that these phosphopeptide- specific antibodies also localize to the centrosome in a cell cycle-dependent fashion. Thus, polyclonal antibodies have been generated against defined phosphopeptides that reiterate many of the immunofluorescence staining properties exhibited by the MPM-2 antibody. These new phosphopeptide-specific antibodies will provide additional probes to examine the phosphorylation of centrosomal components and the functional consequences of their phosphorylation during mitosis. (C) 2000 Wiley-Liss, Inc.
UR - http://www.scopus.com/inward/record.url?scp=0034212756&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034212756&partnerID=8YFLogxK
U2 - 10.1002/(SICI)1097-0029(20000601)49:5<458::AID-JEMT8>3.3.CO;2-R
DO - 10.1002/(SICI)1097-0029(20000601)49:5<458::AID-JEMT8>3.3.CO;2-R
M3 - Article
C2 - 10842373
AN - SCOPUS:0034212756
SN - 1059-910X
VL - 49
SP - 458
EP - 466
JO - Microscopy Research and Technique
JF - Microscopy Research and Technique
IS - 5
ER -