Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain

Christopher J. Hogan, Harrison Wein, Linda Wordeman, Jonathan M. Scholey, Kenneth E. Sawin, W. Zacheus Cande

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Isolated central spindles or spindles in detergent-permeabilized cells from the diatom Cylindrotheca fusiformis can undergo ATP-dependent reactivation of spindle elongation in vitro. We have used a peptide antibody raised against a 10-amino acid portion common to the kinesin superfamily motor domain to look for kinesin-like motor activity during anaphase B of mitosis. The peptide antibody localizes to central spindles. Upon ATP reactivation of spindle elongation, antigens recognized by the antibody are associated exclusively with the central spindle midzone where antiparallel microtubules of each half-spindle overlap. The antibody recognizes several polypeptides by immunoblot using isolated spindle extracts. One of these polypeptides behaves like kinesin with respect to nucleotide-specific binding to and release from taxol-stabilized microtubules. Preincubation of the spindle model with the peptide antibody inhibits subsequent ATP reactivation of spindle elongation. Coincubation of the peptide antibody with peptide antigen rescues spindle function. These results support a role for kinesin-related protein(s) in spindle elongation (anaphase B) of mitosis and suggest that one or several polypeptides that we have identified in spindle extracts may fulfill this function.

Original languageEnglish (US)
Pages (from-to)6611-6615
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number14
StatePublished - Jul 15 1993

Fingerprint

Kinesin
Anaphase
Peptides
Antibodies
Adenosine Triphosphate
Mitosis
Microtubules
Antigens
Diatoms
In Vitro Techniques
Inhibition (Psychology)
Paclitaxel
Detergents
Motor Activity
Nucleotides
Amino Acids

Keywords

  • Anaphase B
  • Kinesin-related proteins
  • Mitosis

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain. / Hogan, Christopher J.; Wein, Harrison; Wordeman, Linda; Scholey, Jonathan M.; Sawin, Kenneth E.; Zacheus Cande, W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 14, 15.07.1993, p. 6611-6615.

Research output: Contribution to journalArticle

Hogan, Christopher J. ; Wein, Harrison ; Wordeman, Linda ; Scholey, Jonathan M. ; Sawin, Kenneth E. ; Zacheus Cande, W. / Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 14. pp. 6611-6615.
@article{6cbcfe07798c4fd28661be62c7512613,
title = "Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain",
abstract = "Isolated central spindles or spindles in detergent-permeabilized cells from the diatom Cylindrotheca fusiformis can undergo ATP-dependent reactivation of spindle elongation in vitro. We have used a peptide antibody raised against a 10-amino acid portion common to the kinesin superfamily motor domain to look for kinesin-like motor activity during anaphase B of mitosis. The peptide antibody localizes to central spindles. Upon ATP reactivation of spindle elongation, antigens recognized by the antibody are associated exclusively with the central spindle midzone where antiparallel microtubules of each half-spindle overlap. The antibody recognizes several polypeptides by immunoblot using isolated spindle extracts. One of these polypeptides behaves like kinesin with respect to nucleotide-specific binding to and release from taxol-stabilized microtubules. Preincubation of the spindle model with the peptide antibody inhibits subsequent ATP reactivation of spindle elongation. Coincubation of the peptide antibody with peptide antigen rescues spindle function. These results support a role for kinesin-related protein(s) in spindle elongation (anaphase B) of mitosis and suggest that one or several polypeptides that we have identified in spindle extracts may fulfill this function.",
keywords = "Anaphase B, Kinesin-related proteins, Mitosis",
author = "Hogan, {Christopher J.} and Harrison Wein and Linda Wordeman and Scholey, {Jonathan M.} and Sawin, {Kenneth E.} and {Zacheus Cande}, W.",
year = "1993",
month = "7",
day = "15",
language = "English (US)",
volume = "90",
pages = "6611--6615",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "14",

}

TY - JOUR

T1 - Inhibition of anaphase spindle elongation in vitro by a peptide antibody that recognizes kinesin motor domain

AU - Hogan, Christopher J.

AU - Wein, Harrison

AU - Wordeman, Linda

AU - Scholey, Jonathan M.

AU - Sawin, Kenneth E.

AU - Zacheus Cande, W.

PY - 1993/7/15

Y1 - 1993/7/15

N2 - Isolated central spindles or spindles in detergent-permeabilized cells from the diatom Cylindrotheca fusiformis can undergo ATP-dependent reactivation of spindle elongation in vitro. We have used a peptide antibody raised against a 10-amino acid portion common to the kinesin superfamily motor domain to look for kinesin-like motor activity during anaphase B of mitosis. The peptide antibody localizes to central spindles. Upon ATP reactivation of spindle elongation, antigens recognized by the antibody are associated exclusively with the central spindle midzone where antiparallel microtubules of each half-spindle overlap. The antibody recognizes several polypeptides by immunoblot using isolated spindle extracts. One of these polypeptides behaves like kinesin with respect to nucleotide-specific binding to and release from taxol-stabilized microtubules. Preincubation of the spindle model with the peptide antibody inhibits subsequent ATP reactivation of spindle elongation. Coincubation of the peptide antibody with peptide antigen rescues spindle function. These results support a role for kinesin-related protein(s) in spindle elongation (anaphase B) of mitosis and suggest that one or several polypeptides that we have identified in spindle extracts may fulfill this function.

AB - Isolated central spindles or spindles in detergent-permeabilized cells from the diatom Cylindrotheca fusiformis can undergo ATP-dependent reactivation of spindle elongation in vitro. We have used a peptide antibody raised against a 10-amino acid portion common to the kinesin superfamily motor domain to look for kinesin-like motor activity during anaphase B of mitosis. The peptide antibody localizes to central spindles. Upon ATP reactivation of spindle elongation, antigens recognized by the antibody are associated exclusively with the central spindle midzone where antiparallel microtubules of each half-spindle overlap. The antibody recognizes several polypeptides by immunoblot using isolated spindle extracts. One of these polypeptides behaves like kinesin with respect to nucleotide-specific binding to and release from taxol-stabilized microtubules. Preincubation of the spindle model with the peptide antibody inhibits subsequent ATP reactivation of spindle elongation. Coincubation of the peptide antibody with peptide antigen rescues spindle function. These results support a role for kinesin-related protein(s) in spindle elongation (anaphase B) of mitosis and suggest that one or several polypeptides that we have identified in spindle extracts may fulfill this function.

KW - Anaphase B

KW - Kinesin-related proteins

KW - Mitosis

UR - http://www.scopus.com/inward/record.url?scp=0027284232&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027284232&partnerID=8YFLogxK

M3 - Article

C2 - 8341676

AN - SCOPUS:0027284232

VL - 90

SP - 6611

EP - 6615

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 14

ER -