Differential localization of the centromere-specific proteins in the major centromeric satellite of Arabidopsis thaliana

Fukashi Shibata, Minoru Murata

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

The 180 bp family of tandem repetitive sequences, which constitutes the major centromeric satellite in Arabidopsis thaliana, is thought to play important roles in kinetochore assembly. To assess the centromere activities of the 180 bp repeats, we performed indirect fluorescence immunolabeling with antibodies against phosphorylated histone H3 at Serl0, HTR12 (Arabidopsis centromeric histone H3 variant) and AtCENP-C (Arabidopsis CENP-C homologue) for the A. thaliana cell cultures. The immunosignals from all three antibodies appeared on all sites of the 180 bp,repeats detected by fluorescence in situ hybridization. However, some of the 180 bp repeat clusters, particularly those that were long or stretched at interphase, were not fully covered with the signals from anti-HTR12 or AtCENP-C. Chromatin fiber immunolabeling clearly revealed that the centromeric proteins examined in this study, localize only at the knobs on the extended chromatin fibers, which form a limited part of the 180 bp clusters. Furthermore, outer HTR12 and inner phosphohistone H3 (Ser1O) localization at the kinetochores of metaphase chromosomes suggests that two kinds of histone H3 (a centromere variant and a phosphorylated form) might be linked to different roles in centromere functionality; the former for spindle-fiber attachment, and the latter for chromatid cohesion.

Original languageEnglish
Pages (from-to)2963-2970
Number of pages8
JournalJournal of Cell Science
Volume117
Issue number14
DOIs
Publication statusPublished - Jun 15 2004

Fingerprint

Centromere
Arabidopsis
Histones
Kinetochores
Chromatin
Proteins
Chromatids
Antibodies
Nucleic Acid Repetitive Sequences
Interphase
Metaphase
Fluorescence In Situ Hybridization
Cell Culture Techniques
Chromosomes
Fluorescence

Keywords

  • 180 bp repeat
  • Arabidopsis thaliana
  • Centromere proteins
  • Histone H3
  • Phosphorylation

ASJC Scopus subject areas

  • Cell Biology

Cite this

Differential localization of the centromere-specific proteins in the major centromeric satellite of Arabidopsis thaliana. / Shibata, Fukashi; Murata, Minoru.

In: Journal of Cell Science, Vol. 117, No. 14, 15.06.2004, p. 2963-2970.

Research output: Contribution to journalArticle

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