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PMID: 17001001 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Formation of functional centromeric chromatin is specified epigenetically in Candida albicans.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 103 ·No. 40 ·2006-10-03 ·Pages 14877-82

Baum M, Sanyal K, Mishra PK, Thaler N, Carbon J

Abstract

In the pathogenic yeast Candida albicans, the 3-kb centromeric DNA regions (CEN) of each of the eight chromosomes have different and unique DNA sequences. The centromeric histone CaCse4p (CENP-A homolog) occurs only within these 3-kb CEN regions to form specialized centromeric chromatin. Centromere activity was maintained on small chromosome fragments derived in vivo by homologous recombination of a native chromosome with linear DNA fragments containing a telomere and a selectable marker. An in vivo derived 85-kb truncated chromosome containing the 3-kb CEN7 locus on 69 kb of chromosome 7 DNA was stably and autonomously maintained in mitosis, indicating that preexisting active CEN chromatin remains functional through many generations. This same 85-kb chromosome fragment, isolated as naked DNA (devoid of chromatin proteins) from C. albicans and reintroduced back into C. albicans cells by standard DNA transformation techniques, was unable to reform functional CEN chromatin and was mitotically unstable. Comparison of active and inactive CEN chromatin digested with micrococcal nuclease revealed that periodic nucleosome arrays are disrupted at active centromeres. Chromatin immunoprecipitation with antibodies against CaCse4p confirmed that CEN7 introduced into C. albicans cells as naked DNA did not recruit CaCse4p or induce its spread to a duplicate region only 7 kb away from active CEN7 chromatin. These results indicate that CaCse4p recruitment and centromere activation are epigenetically specified and maintained in C. albicans.

MeSH Terms
Base Pairing Candida albicans/genetics Centromere/metabolism Chromosomal Proteins, Non-Histone/metabolism Chromosomes, Fungal/genetics DNA Packaging/genetics DNA, Fungal/metabolism Epigenesis, Genetic Fungal Proteins/metabolism Histones/metabolism Micrococcal Nuclease/metabolism Mitosis/physiology Molecular Sequence Data Nucleosomes/metabolism Plasmids/metabolism
Chemicals
Chromosomal Proteins, Non-Histone Cse4 protein, Candida albicans DNA, Fungal Fungal Proteins Histones Nucleosomes Micrococcal Nuclease
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Baum Mary
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA 93106-9610, USA.
Sanyal Kaustuv
Mishra Prashant K
Thaler Nathaniel
Carbon John
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0606958103
Published
2006-10-03
Epub
2006-00-26
Pages
14877-82
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1595444
Subset
IM
Grants
NCI NIH HHS · R01 CA011034 · United States
NCI NIH HHS · CA 11034 · United States
Databases
GENBANK
DQ349190, DQ349191, DQ349192, DQ349193, DQ349194, DQ349195, DQ349196, DQ349197, DQ349198, DQ349199, DQ349200, DQ349201
External Links
PubMed source DOI: 10.1073/pnas.0606958103 PMC Full Text View journal details
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