Kinetochores Prevent Repair of UV Damage in <i>Saccharomyces cerevisiae</i> Centromeres

Capiaghi, Christoph; Thoma, Fritz

doi:10.1128/mcb.24.16.6907-6918.2004

Cited by 14 publications

(7 citation statements)

References 72 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…No repair was detected in the centromere of chromosome XIV, whereas the centromere flanking regions were efficiently repaired (Figure 2). This supports our previous results that showed a strong inhibition of photoreactivation and nucleotide excision repair by kinetochore proteins (Capiaghi et al , 2004).…”

Section: Resultssupporting

confidence: 93%

“…CPDs were mapped by indirect end labeling and quantified as described (Suter et al , 1997; Capiaghi et al , 2004). This method allows correlating CPD distribution with positioned nucleosomes, linker DNA, and nuclease‐sensitive regions (compare Figures 1, 2 and 3).…”

Section: Methodsmentioning

confidence: 99%

“…Chromatin isolation and digestion with micrococcal nuclease digestion was carried out as described (Capiaghi et al , 2004). The cutting sites were analyzed by indirect end labeling as described for mapping CPDs, except that a neutral agarose gel was used for Figure 3A.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Rapid accessibility of nucleosomal DNA in yeast on a second time scale

Bucceri

Kapitza

Thoma

2006

EMBO J

Self Cite

View full text Add to dashboard Cite

Packaging DNA in nucleosomes and higher-order chromatin structures restricts its accessibility and constitutes a barrier for all DNA transactions including gene regulation and DNA repair. How and how fast proteins find access to DNA buried in chromatin of living cells is poorly understood. To address this question in a real time in vivo approach, we investigated DNA repair by photolyase in yeast. We show that overexpressed photolyase, a lightdependent DNA-repair enzyme, recognizes and repairs UVdamaged DNA within seconds. Rapid repair was observed in various nucleosomal regions of the genome including inactive and active genes and repressed promoters. About 50% of cyclobutane pyrimidine dimers were removed in 5 s, 480% in 90 s. Heterochromatin was repaired within minutes, centromeres were not repaired. Consistent with fast conformational transitions of nucleosomes observed in vitro, this rapid repair strongly suggests that spontaneous unwrapping of nucleosomes rather than histone dissociation or chromatin remodeling provides DNA access. The data impact our view on the repressive and dynamic nature of chromatin and illustrate how proteins like photolyase can access DNA in structurally and functionally diverse chromatin regions.

show abstract

Section: Resultssupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Rapid accessibility of nucleosomal DNA in yeast on a second time scale

Bucceri

Kapitza

Thoma

2006

EMBO J

Self Cite

View full text Add to dashboard Cite

show abstract

“…We also examined the histone acetylation level (histones H3 and H4) in the centromere sequence, CEN14. The CEN14 sequence is packed in a compact chromatin structure termed the kinetochore, where UV-induced DNA damage is heavily resistant to NER (26). The histone acetylation level at CEN14 was constant before and after UV irradiation and served as an internal control for the examination of the histone acetylation level at MFA2.…”

Section: Gcn5p Is Required For Efficient Ner Of Uv-induced Cpds In Thementioning

confidence: 99%

UV irradiation stimulates histone acetylation and chromatin remodeling at a repressed yeast locus

Teng

Liu

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

136

190

View full text Add to dashboard Cite

Chromatin immunoprecipitation with anti-acetyl histone H3 (K9 and K14) and anti-acetyl histone H4 (K5, K8, K12, and K16) antibodies shows that Lys-9 and͞or Lys-14 of histone H3, but not the relevant sites of histone H4 in nucleosomes at the repressed MFA2 promoter, are hyperacetylated after UV irradiation. This level of histone hyperacetylation diminishes gradually as repair proceeds. Accompanying this, chromatin in the promoter becomes more accessible to restriction enzymes after UV irradiation and returns to the pre-UV state gradually. UV-related histone hyperacetylation and chromatin remodeling in the MFA2 promoter depend on Gcn5p and partially on Swi2p, respectively. Deletion of GCN5, but not of SWI2, impairs repair of DNA damage in the MFA2 promoter. The post-UV histone modifications and chromatin remodeling at the repressed MFA2 promoter do not activate MFA2 transcriptionally, nor do they require damage recognition by Rad4p or Rad14p. Furthermore, we show that UV irradiation triggers genome-wide histone hyperacetylation at both histone H3 and H4. These experiments indicate that chromatin at a yeast repressed locus undergoes active change after UV radiation treatment and that failure to achieve histone H3 hyperacetylation impairs the repair of DNA damage.nucleotide ͉ excision repair ͉ Saccharomyces cerevisiae P ackaging DNA into chromatin constrains the genome into the cell nucleus and plays important roles in DNA metabolism. The dynamics of chromatin are finely regulated to control DNA function in response to various stimuli. During activation of gene expression the binding of transcriptional activators and coactivators to promoters results in perturbation of repressive chromatin in the promoter. These events facilitate access of subsequent incoming transcriptional factors to DNA. The changes in repressive chromatin often include chromatin remodeling by various complexes, e.g., SWI͞SNF, and acetylation of histones by histone acetyltransferases, e.g., Gcn5p (1, 2).The regulatory roles of histone acetylation and chromatin remodeling, although well documented, are largely confined to transcription initiation. Their roles in other events such as nucleotide excision repair (NER) that also operates on a chromatin template are only beginning to be explored. NER is a DNA repair pathway with Ͼ30 proteins involved in removing damage from naked DNA (3, 4). Studies have shown that the overall repair of DNA damage by NER is less efficient in reconstituted nucleosomes than in naked DNA (5-7), giving an indication that nucleosomes on damaged DNA inhibit efficient NER. In vivo, early studies with human cells showed that increasing the histone acetylation level overall in chromatin by inhibiting histone deacetylase activities enhances repair synthesis during NER (8-12). Chromatin rearrangement during repair synthesis in NER was also observed as newly repaired DNA in human fibroblasts is more sensitive to nuclease than bulk DNA in chromatin. The nascent repair patch lacks a canonical nucleosome DNase I footprinting, and thi...

show abstract

“…Already, Thoma has shown that NER is inefficient in certain sequences in the centromeric regions. 17 Some chromatin remodeling must occur in relation to DNA replication. Does repair of these regions come under the auspices of certain remodeling events related to replication and the cell cycle rather than transcription?…”

Section: Histone Acetylation Chromatin Remodeling and Nucleotide Excmentioning

confidence: 99%

Histone Acetylation, Chromatin Remodelling and Nucleotide Excision Repair: Hint from the Study on MFA2 in Saccharomyces cerevisiae

Waters²

2005

Cell Cycle

View full text Add to dashboard Cite

Nucleotide excision repair (NER) is a sophisticated repair pathway that the cell utilizes to remove a broad range of DNA damage to help maintain the functional integrity of the genome. In the context of DNA packaged into chromosomes it is clear that the NER machinery does not repair all regions with equal efficiency. Recently, we found after UV that histone acetylation and chromatin remodelling were activated. UV irradiation triggers genome-wide histone hyperacetylation at both histone H3 and H4. However, in nucleosomes at the repressed MFA2 promoter only histone H3, but not histone H4, is hyperacetylated following UV. This Gcn5p-mediated histone H3 hyperacetylation enables efficient NER at MFA2. Chromatin in this promoter also becomes more accessible after UV. This is not dependent on Gcn5p, yet it is partially dependent on Swi2p. In later repair times both events gradually return to the pre-UV state. The post-UV histone modifications and chromatin remodelling at the repressed MFA2 promoter do not activate MFA2 transcription, nor do they require damage recognition by Rad4p or Rad14p. These experiments indicate early events are triggered in chromatin in response to UV treatment, and they are likely needed for efficient NER.

show abstract

Kinetochores Prevent Repair of UV Damage in Saccharomyces cerevisiae Centromeres

Cited by 14 publications

References 72 publications

Rapid accessibility of nucleosomal DNA in yeast on a second time scale

Rapid accessibility of nucleosomal DNA in yeast on a second time scale

UV irradiation stimulates histone acetylation and chromatin remodeling at a repressed yeast locus

Histone Acetylation, Chromatin Remodelling and Nucleotide Excision Repair: Hint from the Study on MFA2 in Saccharomyces cerevisiae

Contact Info

Product

Resources

About