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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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Genetic screen for modifiers of His1-dependent adult viability using mis-expression alleles on the second chromosome of Drosophila melanogaster. (A) A genetic cross for screening homozygous viable and fertile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/P{EP} males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). Positions of Act5C-GAL4, pINT-H14M and P{EP} insertions are indicated by black, gray, and white boxes, respectively. The endogenous gene affected by the UAS promoter in the P{EP} insertion is indicated by a hatched box. (B) A genetic cross for screening homozygous lethal and male sterile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/SM5 or P{EP}/CyO males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). (C) Over-represented GO terms and their members among alleles genetically interacting with His1. Gene symbols representing enhancers and suppressors are indicated by red and blue type, respectively. P values are calculated by the hypergeometric test.
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fig1: Genetic screen for modifiers of His1-dependent adult viability using mis-expression alleles on the second chromosome of Drosophila melanogaster. (A) A genetic cross for screening homozygous viable and fertile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/P{EP} males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). Positions of Act5C-GAL4, pINT-H14M and P{EP} insertions are indicated by black, gray, and white boxes, respectively. The endogenous gene affected by the UAS promoter in the P{EP} insertion is indicated by a hatched box. (B) A genetic cross for screening homozygous lethal and male sterile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/SM5 or P{EP}/CyO males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). (C) Over-represented GO terms and their members among alleles genetically interacting with His1. Gene symbols representing enhancers and suppressors are indicated by red and blue type, respectively. P values are calculated by the hypergeometric test.

Mentions: Ten virgin Act5C-GAL4, pINT-H14M/SM5 or Act5C-GAL4/SM5 females were mated with 10 males carrying an EP insertion on the second chromosome (534 alleles), either homozygous or balanced heterozygous (Figure 1, A and B), and reared at 27°. Otherwise, the crosses were performed, and the adult progeny were scored exactly as described above. At least 50 adult F1 flies were scored for each cross.


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)

Genetic screen for modifiers of His1-dependent adult viability using mis-expression alleles on the second chromosome of Drosophila melanogaster. (A) A genetic cross for screening homozygous viable and fertile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/P{EP} males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). Positions of Act5C-GAL4, pINT-H14M and P{EP} insertions are indicated by black, gray, and white boxes, respectively. The endogenous gene affected by the UAS promoter in the P{EP} insertion is indicated by a hatched box. (B) A genetic cross for screening homozygous lethal and male sterile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/SM5 or P{EP}/CyO males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). (C) Over-represented GO terms and their members among alleles genetically interacting with His1. Gene symbols representing enhancers and suppressors are indicated by red and blue type, respectively. P values are calculated by the hypergeometric test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390582&req=5

fig1: Genetic screen for modifiers of His1-dependent adult viability using mis-expression alleles on the second chromosome of Drosophila melanogaster. (A) A genetic cross for screening homozygous viable and fertile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/P{EP} males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). Positions of Act5C-GAL4, pINT-H14M and P{EP} insertions are indicated by black, gray, and white boxes, respectively. The endogenous gene affected by the UAS promoter in the P{EP} insertion is indicated by a hatched box. (B) A genetic cross for screening homozygous lethal and male sterile EP insertions on the second chromosome. Heterozygous Act5C-GAL4, pINT-H14M/SM5 females were mated to P{EP}/SM5 or P{EP}/CyO males, and the ratios of H1-depleted and normal adult progeny were scored based on the balancer phenotypic marker (Cy). (C) Over-represented GO terms and their members among alleles genetically interacting with His1. Gene symbols representing enhancers and suppressors are indicated by red and blue type, respectively. P values are calculated by the hypergeometric test.
Mentions: Ten virgin Act5C-GAL4, pINT-H14M/SM5 or Act5C-GAL4/SM5 females were mated with 10 males carrying an EP insertion on the second chromosome (534 alleles), either homozygous or balanced heterozygous (Figure 1, A and B), and reared at 27°. Otherwise, the crosses were performed, and the adult progeny were scored exactly as described above. At least 50 adult F1 flies were scored for each cross.

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