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A correction to this article has been published: J. Cell Biol. 155 (1) 167
Published 20 August 2001. doi:10.1083/jcb.200103001
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© The Rockefeller University Press, 0021-9525/2001/8/683 $5.00
The Journal of Cell Biology, Volume 154, Number 4, August 20, 2001 683-690

Report

Centromere identity in Drosophila is not determined in vivo by replication timing



Beth Sullivan and Gary Karpen

Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037

Address correspondence to Gary Karpen, MCBL, Salk Institute, 10010 N. Torrey Pines Rd., La Jolla, CA 92037. Tel.: (858) 453-4100, ext. 1473. Fax: (858) 622-0417. E-mail: karpen{at}salk.edu

Centromeric chromatin is uniquely marked by the centromere-specific histone CENP-A. For assembly of CENP-A into nucleosomes to occur without competition from H3 deposition, it was proposed that centromeres are among the first or last sequences to be replicated. In this study, centromere replication in Drosophila was studied in cell lines and in larval tissues that contain minichromosomes that have structurally defined centromeres. Two different nucleotide incorporation methods were used to evaluate replication timing of chromatin containing CID, a Drosophila homologue of CENP-A. Centromeres in Drosophila cell lines were replicated throughout S phase but primarily in mid S phase. However, endogenous centromeres and X-derived minichromosome centromeres in vivo were replicated asynchronously in mid to late S phase. Minichromosomes with structurally intact centromeres were replicated in late S phase, and those in which centric and surrounding heterochromatin were partially or fully deleted were replicated earlier in mid S phase. We provide the first in vivo evidence that centromeric chromatin is replicated at different times in S phase. These studies indicate that incorporation of CID/CENP-A into newly duplicated centromeres is independent of replication timing and argue against determination of centromere identity by temporal sequestration of centromeric chromatin replication relative to bulk genomic chromatin.

Key Words: kinetochore; CENP-A/CID; centromere; replication; neocentromere


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