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PMID: 8909532 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Dynamic elastic behavior of alpha-satellite DNA domains visualized in situ in living human cells.

The Journal of cell biology ·Vol. 135 ·No. 3 ·1996-11-00 ·Pages 545-57

Shelby RD, Hahn KM, Sullivan KF

Abstract

We have constructed a fluorescent alpha-satellite DNA-binding protein to explore the motile and mechanical properties of human centromeres. A fusion protein consisting of human CENP-B coupled to the green fluorescent protein (GFP) of A. victoria specifically targets to centromeres when expressed in human cells. Morphometric analysis revealed that the alpha-satellite DNA domain bound by CENPB-GFP becomes elongated in mitosis in a microtubule-dependent fashion. Time lapse confocal microscopy in live mitotic cells revealed apparent elastic deformations of the central domain of the centromere that occurred during metaphase chromosome oscillations. These observations demonstrate that the interior region of the centromere behaves as an elastic element that could play a role in the mechanoregulatory mechanisms recently identified at centromeres. Fluorescent labeling of centromeres revealed that they disperse throughout the nucleus in a nearly isometric expansion during chromosome decondensation in telophase and early G1. During interphase, centromeres were primarily stationary, although motility of individual or small groups of centromeres was occasionally observed at very slow rates of 7-10 microns/h.

MeSH Terms
Autoantigens Cell Line Centromere Centromere Protein B Chromatin Chromosomal Proteins, Non-Histone DNA, Satellite/analysis DNA-Binding Proteins Fluorescent Dyes Green Fluorescent Proteins HeLa Cells Humans Interphase Luminescent Proteins Microscopy, Confocal/methods Microscopy, Fluorescence Microtubules Mitosis Recombinant Fusion Proteins
Chemicals
Autoantigens CENPB protein, human Centromere Protein B Chromatin Chromosomal Proteins, Non-Histone DNA, Satellite DNA-Binding Proteins Fluorescent Dyes Luminescent Proteins Recombinant Fusion Proteins Green Fluorescent Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Shelby R D
Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037, USA.
Hahn K M
Sullivan K F
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.3.545
Published
1996-11-00
Pages
545-57
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2121065
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.135.3.545 PMC Full Text View journal details
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