Different roles of histone H3 lysine 4 methylation in chromatin maintenance

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Abstract

Histone H3 methyltransferases are involved in the epigenetic control of transcription and heterochromatin maintenance. In Saccharomyces cerevisiae, deletion of a histone H3 methyltransferase SET1 leads to the induction of a subset of stress responsive genes in a Rad53 dependent manner. This type of induction was observed only in the absence of SET1 and not in the absence of other histone methyltransferases, SET2 or DOT1. We show that the increased expression of the stress responsive genes results from a lack of histone H3 lysine (K) 4 methylation. The loss of mono-methylation on H3 K4 is necessary to increase the expression of the stress responsive genes, while the loss of di- or tri-methylation induced by deletion of either RRM domain of Set1 or the upstream effector molecules hardly affected their expression. These results suggest that mono- and multiple methylation of H3 K4 have different roles. The mono-methylation of H3 K4 might be required for the global integrity of chromatin structure, which is normally monitored by the Rad53 dependent chromatin surveillance system.

Section snippets

Materials and methods

Yeast strains and plasmids. The yeast strains used in this study are summarized in Table 1. The histone H3 mutants were constructed by shuffling each plasmid containing histone H3 (WT or mutant alleles) and H4 into WZY42 (from Dent-Roth) to make YC85 (wild-type, pWZ414-F13), YC86 (pRS314-H3(K4A)-H4), YC87 (pRS314-H3(K36A)-H4), YC77 (pRS314-H3(K79A)-H4), and YC79 (pRS314-H3(K4A/K79A)-H4). Wild-type, K4A, and K36A mutant plasmids were gifts from Buratowski. An immunoblotting assay was performed

The expression of stress responsive genes is specific to the SET1 deletion

Deletion of SET1 increases the DNA repair capacities of the checkpoint mutants by inducing repair genes via Rad53 [25]. However, it is so far unclear which SET1 deletion is required for triggering the RAD53 dependent pathway. Possible explanations for this feature of SET1 deletion include loss of Set1 protein (regardless of HMT activity), loss of histone H3 K4 methylation, or loss of methylation of an unknown factor(s). As a first step toward clarifying this, the expression of several stress

Discussion

This study showed that set1Δ resulted in the induction of several stress responsive genes in a RAD53 dependent manner. Among the methylation targets of Set1, histone H3 K4 might be directly involved in this pathway, because a loss of histone H3 K4 methylation is necessary to induce the target genes. Interestingly, this pathway has differentiated the roles of H3 K4 methylation states. The loss of di- or tri-methylation, which is implicated in the active transcription, did not induce the stress

Acknowledgments

We thank Drs. S. Buratowski, A. Verreault, T. Kouzarides, S. Roeder, S. Dent-Roth, S.D. Briggs., S. Hanes, and B. Cairns for yeast strains and plasmids and A. Verreault for critical reading of the manuscript. This work was supported by the grant from the Korea Research Foundation Grant (R04-2004-000-10188-0, 2005-0497-000) to E.-J. Cho.

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