Genome-wide analysis of regulation of gene expression and H3K9me2 distribution by JIL-1 kinase mediated histone H3S10 phosphorylation in Drosophila

In this study we have determined the genome-wide relationship of JIL-1 kinase mediated H3S10 phosphorylation with gene expression and the distribution of the epigenetic H3K9me2 mark. We show in wild-type salivary gland cells that the H3S10ph mark is predominantly enriched at active genes whereas the H3K9me2 mark is largely associated with inactive genes. Comparison of global transcription profiles in salivary glands from wild-type and JIL-1 null mutant larvae revealed that the expression levels of 1539 genes changed at least 2-fold in the mutant and that a substantial number (49%) of these genes were upregulated whereas 51% were downregulated. Furthermore, the results showed that downregulation of genes in the mutant was correlated with higher levels or acquisition of the H3K9me2 mark whereas upregulation of a gene was correlated with loss of or diminished H3K9 dimethylation. These results are compatible with a model where gene expression levels are modulated by the levels of the H3K9me2 mark independent of the state of the H3S10ph mark, which is not required for either transcription or gene activation to occur. Rather, H3S10 phosphorylation functions to indirectly maintain active transcription by counteracting H3K9 dimethylation and gene silencing.

[1]  Tobias Straub,et al.  Global Analysis of the Relationship between JIL-1 Kinase and Transcription , 2011, PLoS genetics.

[2]  Yang Zhang Progress and challenges in protein structure prediction. , 2008, Current opinion in structural biology.

[3]  K. Johansen,et al.  Genetic and phenotypic analysis of alleles of the Drosophila chromosomal JIL-1 kinase reveals a functional requirement at multiple developmental stages. , 2003, Genetics.

[4]  Xiaomin Bao,et al.  The JIL-1 histone H3S10 kinase regulates dimethyl H3K9 modifications and heterochromatic spreading in Drosophila , 2005, Development.

[5]  K. Johansen,et al.  The JIL-1 Tandem Kinase Mediates Histone H3 Phosphorylation and Is Required for Maintenance of Chromatin Structure in Drosophila , 2001, Cell.

[6]  K. Johansen,et al.  Jil-1, a Chromosomal Kinase Implicated in Regulation of Chromatin Structure, Associates with the Male Specific Lethal (Msl) Dosage Compensation Complex , 2000, The Journal of cell biology.

[7]  Hadley Wickham,et al.  ggplot2 - Elegant Graphics for Data Analysis (2nd Edition) , 2017 .

[8]  Serban Nacu,et al.  Fast and SNP-tolerant detection of complex variants and splicing in short reads , 2010, Bioinform..

[9]  K M Johansen,et al.  JIL-1: a novel chromosomal tandem kinase implicated in transcriptional regulation in Drosophila. , 1999, Molecular cell.

[10]  Xiaomin Bao,et al.  The chromodomain protein, Chromator, interacts with JIL-1 kinase and regulates the structure of Drosophila polytene chromosomes , 2006, Journal of Cell Science.

[11]  Sanzhen Liu,et al.  Gene Mapping via Bulked Segregant RNA-Seq (BSR-Seq) , 2012, PloS one.

[12]  Robert Krencik,et al.  The COOH-terminal Domain of the JIL-1 Histone H3S10 Kinase Interacts with Histone H3 and Is Required for Correct Targeting to Chromatin* , 2008, Journal of Biological Chemistry.

[13]  K. Johansen,et al.  Regulation of chromatin structure by histone H3S10 phosphorylation , 2006, Chromosome Research.

[14]  K. Johansen,et al.  H3S10 phosphorylation by the JIL-1 kinase regulates H3K9 dimethylation and gene expression at the white locus in Drosophila , 2012, Fly.

[15]  Aaron R. Quinlan,et al.  BIOINFORMATICS APPLICATIONS NOTE , 2022 .

[16]  Yang Zhang,et al.  I-TASSER: a unified platform for automated protein structure and function prediction , 2010, Nature Protocols.

[17]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[18]  K. Johansen,et al.  The epigenetic H3S10 phosphorylation mark is required for counteracting heterochromatic spreading and gene silencing in Drosophila melanogaster , 2011, Journal of Cell Science.

[19]  K. Johansen,et al.  Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling , 2010, Journal of visualized experiments : JoVE.

[20]  K. Mechtler,et al.  14‐3‐3 Proteins recognize a histone code at histone H3 and are required for transcriptional activation , 2008, The EMBO journal.

[21]  C. S. Karam,et al.  Phosphorylation of histone H3 at Ser10 facilitates RNA polymerase II release from promoter-proximal pausing in Drosophila. , 2007, Genes & development.

[22]  C. S. Karam,et al.  14-3-3 Mediates Histone Cross-Talk during Transcription Elongation in Drosophila , 2010, PLoS genetics.

[23]  Clifford A. Meyer,et al.  Model-based Analysis of ChIP-Seq (MACS) , 2008, Genome Biology.

[24]  Chen Zeng,et al.  A clustering approach for identification of enriched domains from histone modification ChIP-Seq data , 2009, Bioinform..

[25]  Changfu Yao,et al.  The chromodomain-containing NH2-terminus of Chromator interacts with histone H1 and is required for correct targeting to chromatin , 2011, Chromosoma.

[26]  M. Gerstein,et al.  Unlocking the secrets of the genome , 2009, Nature.

[27]  Sandrine Dudoit,et al.  Evaluation of statistical methods for normalization and differential expression in mRNA-Seq experiments , 2010, BMC Bioinformatics.

[28]  Xiaomin Bao,et al.  Loss-Of-Function Alleles of the JIL-1 Kinase Are Strong Suppressors of Position Effect Variegation of the wm4 Allele in Drosophila , 2006, Genetics.

[29]  David S. Lapointe,et al.  ChIPpeakAnno: a Bioconductor package to annotate ChIP-seq and ChIP-chip data , 2010, BMC Bioinformatics.

[30]  K. Johansen,et al.  The JIL-1 kinase regulates the structure of Drosophila polytene chromosomes , 2005, Chromosoma.

[31]  Salvatore Oliviero,et al.  Histone Crosstalk between H3S10ph and H4K16ac Generates a Histone Code that Mediates Transcription Elongation , 2009, Cell.

[32]  Changfu Yao,et al.  Evidence against a Role for the JIL-1 Kinase in H3S28 Phosphorylation and 14-3-3 Recruitment to Active Genes in Drosophila , 2013, PloS one.

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

[34]  K. Johansen,et al.  Chromator, a novel and essential chromodomain protein interacts directly with the putative spindle matrix protein skeletor , 2004, Journal of cellular biochemistry.

[35]  Steven P Lund,et al.  Statistical Applications in Genetics and Molecular Biology Detecting Differential Expression in RNA-sequence Data Using Quasi-likelihood with Shrunken Dispersion Estimates , 2012 .

[36]  K. Johansen,et al.  JIL-1 and Su(var)3-7 Interact Genetically and Counteract Each Other's Effect on Position-Effect Variegation in Drosophila , 2010, Genetics.

[37]  K. Johansen,et al.  A Balance Between Euchromatic (JIL-1) and Heterochromatic [SU(VAR)2-5 and SU(VAR)3-9] Factors Regulates Position-Effect Variegation in Drosophila , 2011, Genetics.

[38]  G. Schotta,et al.  Su(var) genes regulate the balance between euchromatin and heterochromatin in Drosophila. , 2004, Genes & development.

[39]  Yang Zhang,et al.  Template‐based modeling and free modeling by I‐TASSER in CASP7 , 2007, Proteins.

[40]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[41]  M. Bownes,et al.  Drosophila: A practical approach: edited by D. B. Roberts IRL Press, 1986. £26.00/$47.00 (xix + 295 pages) ISBN 0 94746 66 7 , 1987 .

[42]  K. Johansen,et al.  Reduced Levels of Su(var)3-9 But Not Su(var)2-5 (HP1) Counteract the Effects on Chromatin Structure and Viability in Loss-of-Function Mutants of the JIL-1 Histone H3S10 Kinase , 2007, Genetics.

[43]  Victor G Corces,et al.  Genome-wide phosphoacetylation of histone H3 at Drosophila enhancers and promoters , 2012, Genome research.

[44]  K. Johansen,et al.  RNA polymerase II-mediated transcription at active loci does not require histone H3S10 phosphorylation in Drosophila , 2008, Development.

[45]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[46]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[47]  M. Lercher,et al.  X-chromosome-wide profiling of MSL-1 distribution and dosage compensation in Drosophila. , 2006, Genes & development.