Comparative analysis of expression of histone H2a genes in mouse

BackgroundAt least 18 replication-dependent histone H2a genes are distributed in 3 Hist gene clusters on different chromosomes of the mouse genome. In this analysis we designed specific PCR primers for each histone H2a transcript and studied the expression levels and patterns using quantitative RT-PCR (qRT-PCR). In addition, we compared histone H3 K9 acetylation levels in the promoter regions of H2a genes by ChIP (chromatin immunoprecipitation) – quantitative PCR (qPCR) analysis.ResultsRT-PCR analysis indicated that all 20 histone H2a genes assessed in this study are expressed. The replication-dependent histone H2a genes have different expression levels but similar expression patterns. Among the 20 histone H2a genes, the expression-level of H2afz, a replication-independent gene, was highest, and that of Hist1h2aa, a replication-dependent gene, was lowest. Among 18 replication-dependent H2a genes, the expression level of Hist3h2a was highest. The ChIP-qPCR analysis showed that histone H3 K9 acetylation levels in promoter regions of both H2afz and Hist3h2a are clearly higher than that in the promoter region of Hist1h2aa. The H3 K9 acetylation level in the promoter of Hist1h2aa is similar to that in the γ-satellite region.ConclusionThese results strongly suggest that histone H3 K9 acetylation plays a role in the expression of histone genes.

[1]  C. Allis,et al.  Histone methylation versus histone acetylation: new insights into epigenetic regulation. , 2001, Current opinion in cell biology.

[2]  M. Osley.  The regulation of histone synthesis in the cell cycle. , 1991, Annual review of biochemistry.

[3]  D. Schümperli,et al.  Regulation of histone mRNA in the unperturbed cell cycle: evidence suggesting control at two posttranscriptional steps , 1991, Molecular and cellular biology.

[4]  Danny Reinberg,et al.  Recent highlights of RNA-polymerase-II-mediated transcription. , 2004, Current opinion in cell biology.

[5]  Hiten D. Madhani,et al.  Conserved Histone Variant H2A.Z Protects Euchromatin from the Ectopic Spread of Silent Heterochromatin , 2003, Cell.

[6]  A. Ehrenhofer-Murray,et al.  Chromatin dynamics at DNA replication, transcription and repair. , 2004, European journal of biochemistry.

[7]  Keji Zhao,et al.  Active chromatin domains are defined by acetylation islands revealed by genome-wide mapping. , 2005, Genes & development.

[8]  Mark Stitt,et al.  Real-time RT-PCR profiling of over 1400 Arabidopsis transcription factors: unprecedented sensitivity reveals novel root- and shoot-specific genes. , 2004, The Plant journal : for cell and molecular biology.

[9]  W. Bonner,et al.  H2A.X. a histone isoprotein with a conserved C-terminal sequence, is encoded by a novel mRNA with both DNA replication type and polyA 3' processing signals. , 1989, Nucleic acids research.

[10]  R. Duronio,et al.  Histone mRNA expression: multiple levels of cell cycle regulation and important developmental consequences. , 2002, Current opinion in cell biology.

[11]  F. Oswald,et al.  The E2F transcription factor activates a replication-dependent human H2A gene in early S phase of the cell cycle , 1996, Molecular and cellular biology.

[12]  William F Marzluff,et al.  The human and mouse replication-dependent histone genes. , 2002, Genomics.

[13]  W. Bonner,et al.  The human histone H2A.Z gene. Sequence and regulation. , 1990, The Journal of biological chemistry.

[14]  Y. Hayashizaki,et al.  A novel replication-independent histone H2a gene in mouse , 2005, BMC Genetics.

[15]  E. Bonnefoy,et al.  Transcription factor YY1 associates with pericentromeric gamma-satellite DNA in cycling but not in quiescent (G0) cells. , 2004, Nucleic acids research.

[16]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[17]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[18]  Y. Hayashizaki,et al.  Naturally occurring antisense RNA of histone H2a in mouse cultured cell lines , 2005, BMC Genetics.