CaltechAUTHORS
Published July 6, 2015 | Version Accepted Version + Supplemental Material
Journal Article Open

Establishment of Centromeric Chromatin by the CENP-A Assembly Factor CAL1 Requires FACT-Mediated Transcription

Creators

  • 1. ROR icon University of Connecticut
  • 2. ROR icon University of Cambridge
  • 3. ROR icon Institute of Biochemistry

Abstract

Centromeres are essential chromosomal structures that mediate accurate chromosome segregation during cell division. Centromeres are specified epigenetically by the heritable incorporation of the centromeric histone H3 variant CENP-A. While many of the primary factors that mediate centromeric deposition of CENP-A are known, the chromatin and DNA requirements of this process have remained elusive. Here, we uncover a role for transcription in Drosophila CENP-A deposition. Using an inducible ectopic centromere system that uncouples CENP-A deposition from endogenous centromere function and cell-cycle progression, we demonstrate that CENP-A assembly by its loading factor, CAL1, requires RNAPII-mediated transcription of the underlying DNA. This transcription depends on the CAL1 binding partner FACT, but not on CENP-A incorporation. Our work establishes RNAPII passage as a key step in chaperone-mediated CENP-A chromatin establishment and propagation.

Additional Information

© 2015 Elsevier. Under an Elsevier user license. Received 7 October 2014, Revised 9 April 2015, Accepted 18 May 2015, Available online 6 July 2015. We thank Bo Reese (CGI) for sequencing support, G. Karpen, S. Hirose, A. Straight, and P. Heun for reagents, I. Cheeseman for critically reading the manuscript, L. Core for suggestions and help with ChIP-chip data analysis, and the Drosophila RNAi Screening Center (DRSC) for resources. This work was supported by awards from the NSF (1024973) (to B.G.M.) and the NIH (GM108829) (to B.G.M.). D.M.G. is supported by grants from Cancer Research UK (C3/A11431) and the Medical Research Council (G1001696); Z.L. is supported by a long-term FEBS Fellowship; and R.J.O. is supported by an NSF award (1244146). Accession Numbers. The accession number for the ChIP-seq and RNA-seq raw data reported in this paper is NCBI: SRP059507.

Attached Files

Accepted Version - nihms693741.pdf

Supplemental Material - 1-s2.0-S1534580715003482-mmc1.pdf

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1-s2.0-S1534580715003482-mmc1.pdf

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Additional details

Identifiers

PMCID
PMC4495351
Eprint ID
105601
DOI
10.1016/j.devcel.2015.05.012
Resolver ID
CaltechAUTHORS:20200928-150624733

Related works

Describes
10.1016/j.devcel.2015.05.012 (DOI)

Funding

NSF
MCB-1024973
NIH
GM108829
Cancer Research UK
C3/A11431
Medical Research Council (UK)
G1001696
Federation of European Biochemical Societies
NSF
MCB-1244146

Dates

Created
2020-09-30
Created from EPrint's datestamp field
Updated
2023-03-16
Created from EPrint's last_modified field