Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase

Chad G. Pearson, Elaine Yeh, Melissa K Gardner, David J Odde, E. D. Salmon, Kerry Bloom

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

With a single microtubule attachment, budding-yeast kinetochores provide an excellent system for understanding the coordinated linkage to dynamic microtubule plus ends for chromosome oscillation and positioning. Fluorescent tagging of kinetochore proteins indicates that, on average, all centromeres are clustered, distinctly separated from their sisters, and positioned equidistant from their respective spindle poles during metaphase. However, individual fluorescent chromosome markers near the centromere transiently reassociate with their sisters and oscillate from one spindle half to the other. To reconcile the apparent disparity between the average centromere position and individual centromere proximal markers, we utilized fluorescence recovery after photobleaching to measure stability of the histone-H3 variant Cse4p/CENP-A. Newly synthesized Cse4p replaces old protein during DNA replication. Once assembled, Cse4-GFP is a physically stable component of centromeres during mitosis. This allowed us to follow centromere dynamics within each spindle half. Kinetochores remain stably attached to dynamic microtubules and exhibit a low incidence of switching orientation or position between the spindle halves. Switching of sister chromatid attachment may be contemporaneous with Cse4p exchange and early kinetochore assembly during S phase; this would promote mixing of chromosome attachment to each spindle pole. Once biorientation is attained, centromeres rarely make excursions beyond their proximal half spindle.

Original languageEnglish (US)
Pages (from-to)1962-1967
Number of pages6
JournalCurrent Biology
Volume14
Issue number21
DOIs
StatePublished - Nov 9 2004

Fingerprint

Kinetochores
kinetochores
Centromere
centromeres
Metaphase
Chromosomes
metaphase
Microtubules
microtubules
Poles
Photobleaching
Spindle Poles
Histones
Yeast
chromosomes
Proteins
Fluorescence
Chromosome Positioning
Recovery
Fluorescence Recovery After Photobleaching

Cite this

Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase. / Pearson, Chad G.; Yeh, Elaine; Gardner, Melissa K; Odde, David J; Salmon, E. D.; Bloom, Kerry.

In: Current Biology, Vol. 14, No. 21, 09.11.2004, p. 1962-1967.

Research output: Contribution to journalArticle

Pearson, Chad G. ; Yeh, Elaine ; Gardner, Melissa K ; Odde, David J ; Salmon, E. D. ; Bloom, Kerry. / Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase. In: Current Biology. 2004 ; Vol. 14, No. 21. pp. 1962-1967.
@article{3d00e80e9d464f598498c12f409197fa,
title = "Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase",
abstract = "With a single microtubule attachment, budding-yeast kinetochores provide an excellent system for understanding the coordinated linkage to dynamic microtubule plus ends for chromosome oscillation and positioning. Fluorescent tagging of kinetochore proteins indicates that, on average, all centromeres are clustered, distinctly separated from their sisters, and positioned equidistant from their respective spindle poles during metaphase. However, individual fluorescent chromosome markers near the centromere transiently reassociate with their sisters and oscillate from one spindle half to the other. To reconcile the apparent disparity between the average centromere position and individual centromere proximal markers, we utilized fluorescence recovery after photobleaching to measure stability of the histone-H3 variant Cse4p/CENP-A. Newly synthesized Cse4p replaces old protein during DNA replication. Once assembled, Cse4-GFP is a physically stable component of centromeres during mitosis. This allowed us to follow centromere dynamics within each spindle half. Kinetochores remain stably attached to dynamic microtubules and exhibit a low incidence of switching orientation or position between the spindle halves. Switching of sister chromatid attachment may be contemporaneous with Cse4p exchange and early kinetochore assembly during S phase; this would promote mixing of chromosome attachment to each spindle pole. Once biorientation is attained, centromeres rarely make excursions beyond their proximal half spindle.",
author = "Pearson, {Chad G.} and Elaine Yeh and Gardner, {Melissa K} and Odde, {David J} and Salmon, {E. D.} and Kerry Bloom",
year = "2004",
month = "11",
day = "9",
doi = "10.1016/j.cub.2004.09.086",
language = "English (US)",
volume = "14",
pages = "1962--1967",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "21",

}

TY - JOUR

T1 - Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase

AU - Pearson, Chad G.

AU - Yeh, Elaine

AU - Gardner, Melissa K

AU - Odde, David J

AU - Salmon, E. D.

AU - Bloom, Kerry

PY - 2004/11/9

Y1 - 2004/11/9

N2 - With a single microtubule attachment, budding-yeast kinetochores provide an excellent system for understanding the coordinated linkage to dynamic microtubule plus ends for chromosome oscillation and positioning. Fluorescent tagging of kinetochore proteins indicates that, on average, all centromeres are clustered, distinctly separated from their sisters, and positioned equidistant from their respective spindle poles during metaphase. However, individual fluorescent chromosome markers near the centromere transiently reassociate with their sisters and oscillate from one spindle half to the other. To reconcile the apparent disparity between the average centromere position and individual centromere proximal markers, we utilized fluorescence recovery after photobleaching to measure stability of the histone-H3 variant Cse4p/CENP-A. Newly synthesized Cse4p replaces old protein during DNA replication. Once assembled, Cse4-GFP is a physically stable component of centromeres during mitosis. This allowed us to follow centromere dynamics within each spindle half. Kinetochores remain stably attached to dynamic microtubules and exhibit a low incidence of switching orientation or position between the spindle halves. Switching of sister chromatid attachment may be contemporaneous with Cse4p exchange and early kinetochore assembly during S phase; this would promote mixing of chromosome attachment to each spindle pole. Once biorientation is attained, centromeres rarely make excursions beyond their proximal half spindle.

AB - With a single microtubule attachment, budding-yeast kinetochores provide an excellent system for understanding the coordinated linkage to dynamic microtubule plus ends for chromosome oscillation and positioning. Fluorescent tagging of kinetochore proteins indicates that, on average, all centromeres are clustered, distinctly separated from their sisters, and positioned equidistant from their respective spindle poles during metaphase. However, individual fluorescent chromosome markers near the centromere transiently reassociate with their sisters and oscillate from one spindle half to the other. To reconcile the apparent disparity between the average centromere position and individual centromere proximal markers, we utilized fluorescence recovery after photobleaching to measure stability of the histone-H3 variant Cse4p/CENP-A. Newly synthesized Cse4p replaces old protein during DNA replication. Once assembled, Cse4-GFP is a physically stable component of centromeres during mitosis. This allowed us to follow centromere dynamics within each spindle half. Kinetochores remain stably attached to dynamic microtubules and exhibit a low incidence of switching orientation or position between the spindle halves. Switching of sister chromatid attachment may be contemporaneous with Cse4p exchange and early kinetochore assembly during S phase; this would promote mixing of chromosome attachment to each spindle pole. Once biorientation is attained, centromeres rarely make excursions beyond their proximal half spindle.

UR - http://www.scopus.com/inward/record.url?scp=7944229979&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=7944229979&partnerID=8YFLogxK

U2 - 10.1016/j.cub.2004.09.086

DO - 10.1016/j.cub.2004.09.086

M3 - Article

VL - 14

SP - 1962

EP - 1967

JO - Current Biology

JF - Current Biology

SN - 0960-9822

IS - 21

ER -