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A genetic screen and transcript profiling reveal a shared regulatory program for Drosophila linker histone H1 and chromatin remodeler CHD1.

Kavi H, Lu X, Xu N, Bartholdy BA, Vershilova E, Skoultchi AI, Fyodorov DV - G3 (Bethesda) (2015)

Bottom Line: It has a profound effect on organization of chromatin into higher-order structures and on recruitment of histone-modifying enzymes to chromatin.We identify 41 mis-expression alleles that enhance and 20 that suppress the effect of His1 depletion in vivo.Specifically, the reduced viability of H1-depleted animals is strongly suppressed by ubiquitous mis-expression of the ATP-dependent chromatin remodeling enzyme CHD1.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461.

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Regulation of immunity and stress response–related genes by H1. (A) Multiple immune response and stress response–related genes are upregulated in H1-depleted larval salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands. Fold changes are calculated as a ratio of signals for H1-depleted samples to those for control samples. Values are normalized to the expression of rp49, are representative of three independent experiments, and are presented in a logarithmic scale. SDs are shown as error bars. Transcripts for Act5C, Gapdh1, and βTub60D are shown as controls. (B) Antimicrobial peptide (AMP) genes are not substantially affected by H1 knockdown in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Transcript for βTub60D is shown as a control. (C) NFκB-family transcription factors Dorsal (dl), Dorsal-related immunity factor (Dif), and Relish (Rel) are not coordinately regulated on H1 depletion in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Black, white, and gray bars indicate transcripts that are significantly upregulated, downregulated, or not affected, respectively, in the H1 knockdown.
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fig2: Regulation of immunity and stress response–related genes by H1. (A) Multiple immune response and stress response–related genes are upregulated in H1-depleted larval salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands. Fold changes are calculated as a ratio of signals for H1-depleted samples to those for control samples. Values are normalized to the expression of rp49, are representative of three independent experiments, and are presented in a logarithmic scale. SDs are shown as error bars. Transcripts for Act5C, Gapdh1, and βTub60D are shown as controls. (B) Antimicrobial peptide (AMP) genes are not substantially affected by H1 knockdown in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Transcript for βTub60D is shown as a control. (C) NFκB-family transcription factors Dorsal (dl), Dorsal-related immunity factor (Dif), and Relish (Rel) are not coordinately regulated on H1 depletion in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Black, white, and gray bars indicate transcripts that are significantly upregulated, downregulated, or not affected, respectively, in the H1 knockdown.

Mentions: To confirm the effects of H1 on these genes, we performed quantitative RT-PCR to compare mRNA expression of some of the immune response–related genes in H1-depleted and control salivary glands. We discovered a consistent correspondence between the microarray and RT-PCR data: genes that were detected as repression targets of H1 by microarray analyses also exhibited upregulation in H1-depleted salivary glands when examined by RT-PCR (Figure 2A and data not shown). Many additional immune response–related genes that were not originally identified as H1 targets by microarray analyses also exhibit strong upregulation (Figure 2A). It is of interest that many of these genes are repressed by a chromatin-remodeling factor CHD1 (Sebald et al. 2012), and Chd1 was identified as one of the His1-interacting genes in our mis-expression screen (Table 2).


A genetic screen and transcript profiling reveal a shared regulatory program for Drosophila linker histone H1 and chromatin remodeler CHD1.

Kavi H, Lu X, Xu N, Bartholdy BA, Vershilova E, Skoultchi AI, Fyodorov DV - G3 (Bethesda) (2015)

Regulation of immunity and stress response–related genes by H1. (A) Multiple immune response and stress response–related genes are upregulated in H1-depleted larval salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands. Fold changes are calculated as a ratio of signals for H1-depleted samples to those for control samples. Values are normalized to the expression of rp49, are representative of three independent experiments, and are presented in a logarithmic scale. SDs are shown as error bars. Transcripts for Act5C, Gapdh1, and βTub60D are shown as controls. (B) Antimicrobial peptide (AMP) genes are not substantially affected by H1 knockdown in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Transcript for βTub60D is shown as a control. (C) NFκB-family transcription factors Dorsal (dl), Dorsal-related immunity factor (Dif), and Relish (Rel) are not coordinately regulated on H1 depletion in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Black, white, and gray bars indicate transcripts that are significantly upregulated, downregulated, or not affected, respectively, in the H1 knockdown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4390582&req=5

fig2: Regulation of immunity and stress response–related genes by H1. (A) Multiple immune response and stress response–related genes are upregulated in H1-depleted larval salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands. Fold changes are calculated as a ratio of signals for H1-depleted samples to those for control samples. Values are normalized to the expression of rp49, are representative of three independent experiments, and are presented in a logarithmic scale. SDs are shown as error bars. Transcripts for Act5C, Gapdh1, and βTub60D are shown as controls. (B) Antimicrobial peptide (AMP) genes are not substantially affected by H1 knockdown in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Transcript for βTub60D is shown as a control. (C) NFκB-family transcription factors Dorsal (dl), Dorsal-related immunity factor (Dif), and Relish (Rel) are not coordinately regulated on H1 depletion in salivary glands. Real-time RT-PCR assays of transcripts were performed in H1-depleted and control salivary glands, analyzed, and presented as in (A). Black, white, and gray bars indicate transcripts that are significantly upregulated, downregulated, or not affected, respectively, in the H1 knockdown.
Mentions: To confirm the effects of H1 on these genes, we performed quantitative RT-PCR to compare mRNA expression of some of the immune response–related genes in H1-depleted and control salivary glands. We discovered a consistent correspondence between the microarray and RT-PCR data: genes that were detected as repression targets of H1 by microarray analyses also exhibited upregulation in H1-depleted salivary glands when examined by RT-PCR (Figure 2A and data not shown). Many additional immune response–related genes that were not originally identified as H1 targets by microarray analyses also exhibit strong upregulation (Figure 2A). It is of interest that many of these genes are repressed by a chromatin-remodeling factor CHD1 (Sebald et al. 2012), and Chd1 was identified as one of the His1-interacting genes in our mis-expression screen (Table 2).

Bottom Line: It has a profound effect on organization of chromatin into higher-order structures and on recruitment of histone-modifying enzymes to chromatin.We identify 41 mis-expression alleles that enhance and 20 that suppress the effect of His1 depletion in vivo.Specifically, the reduced viability of H1-depleted animals is strongly suppressed by ubiquitous mis-expression of the ATP-dependent chromatin remodeling enzyme CHD1.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461.

Show MeSH
Related in: MedlinePlus