Neocentromeres form efficiently at multiple possible loci in candida albicans

Carrie Ketel, Helen S W Wang, Mark McClellan, Kelly Bouchonville, Anna Selmecki, Tamar Lahav, Maryam Gerami-Nejad, Judith Berman

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: ''proximal neoCEN'' and ''distal neoCEN'' strains. Neocentromeres in the distal neoCEN strains formed at loci about 200-450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere. Copyright:

Original languageEnglish
JournalPLoS Genetics
Volume5
Issue number3
DOIs
StatePublished - Mar 2009

Fingerprint

Centromere
centromeres
Candida albicans
chromosomes
DNA
chromosome
loci
Chromosomes
Kinetochores
kinetochores
Heterochromatin
heterochromatin
Genes
gene
nucleotide sequences
High-Throughput Nucleotide Sequencing
Chromosomal Instability
Chromosomes, Human, Pair 5
genes
Chromatin Immunoprecipitation

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology
  • Ecology, Evolution, Behavior and Systematics
  • Cancer Research
  • Genetics(clinical)

Cite this

Ketel, C., Wang, H. S. W., McClellan, M., Bouchonville, K., Selmecki, A., Lahav, T., ... Berman, J. (2009). Neocentromeres form efficiently at multiple possible loci in candida albicans. PLoS Genetics, 5(3). https://doi.org/10.1371/journal.pgen.1000400

Neocentromeres form efficiently at multiple possible loci in candida albicans. / Ketel, Carrie; Wang, Helen S W; McClellan, Mark; Bouchonville, Kelly; Selmecki, Anna; Lahav, Tamar; Gerami-Nejad, Maryam; Berman, Judith.

In: PLoS Genetics, Vol. 5, No. 3, 03.2009.

Research output: Contribution to journalArticle

Ketel, C, Wang, HSW, McClellan, M, Bouchonville, K, Selmecki, A, Lahav, T, Gerami-Nejad, M & Berman, J 2009, 'Neocentromeres form efficiently at multiple possible loci in candida albicans', PLoS Genetics, vol. 5, no. 3. https://doi.org/10.1371/journal.pgen.1000400
Ketel, Carrie ; Wang, Helen S W ; McClellan, Mark ; Bouchonville, Kelly ; Selmecki, Anna ; Lahav, Tamar ; Gerami-Nejad, Maryam ; Berman, Judith. / Neocentromeres form efficiently at multiple possible loci in candida albicans. In: PLoS Genetics. 2009 ; Vol. 5, No. 3.
@article{f11d452e020342b8b922ad86c9ea4f16,
title = "Neocentromeres form efficiently at multiple possible loci in candida albicans",
abstract = "Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: ''proximal neoCEN'' and ''distal neoCEN'' strains. Neocentromeres in the distal neoCEN strains formed at loci about 200-450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere. Copyright:",
author = "Carrie Ketel and Wang, {Helen S W} and Mark McClellan and Kelly Bouchonville and Anna Selmecki and Tamar Lahav and Maryam Gerami-Nejad and Judith Berman",
year = "2009",
month = "3",
doi = "10.1371/journal.pgen.1000400",
language = "English",
volume = "5",
journal = "PLoS Genetics",
issn = "1553-7390",
publisher = "Public Library of Science",
number = "3",

}

TY - JOUR

T1 - Neocentromeres form efficiently at multiple possible loci in candida albicans

AU - Ketel, Carrie

AU - Wang, Helen S W

AU - McClellan, Mark

AU - Bouchonville, Kelly

AU - Selmecki, Anna

AU - Lahav, Tamar

AU - Gerami-Nejad, Maryam

AU - Berman, Judith

PY - 2009/3

Y1 - 2009/3

N2 - Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: ''proximal neoCEN'' and ''distal neoCEN'' strains. Neocentromeres in the distal neoCEN strains formed at loci about 200-450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere. Copyright:

AB - Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: ''proximal neoCEN'' and ''distal neoCEN'' strains. Neocentromeres in the distal neoCEN strains formed at loci about 200-450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere. Copyright:

UR - http://www.scopus.com/inward/record.url?scp=62149122605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=62149122605&partnerID=8YFLogxK

U2 - 10.1371/journal.pgen.1000400

DO - 10.1371/journal.pgen.1000400

M3 - Article

C2 - 19266018

AN - SCOPUS:62149122605

VL - 5

JO - PLoS Genetics

JF - PLoS Genetics

SN - 1553-7390

IS - 3

ER -