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Recruitment of the ATP-dependent chromatin remodeler dMi-2 to the transcribed region of active heat shock genes.

Mathieu EL, Finkernagel F, Murawska M, Scharfe M, Jarek M, Brehm A - Nucleic Acids Res. (2012)

Bottom Line: Interestingly, dMi-2 binding extends several hundred base pairs beyond the polyadenylation site into the region where transcriptional termination occurs.We find that dMi-2 does not associate with the entire nucleosome-depleted hsp70 locus 87A.Rather, dMi-2 binding is restricted to transcribed regions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Molecular Biology and Tumor Research, Philipps-University, Emil-Mannkopff-Strasse 2, 35037 Marburg, Germany.

ABSTRACT
The ATP-dependent chromatin remodeler dMi-2 can play both positive and negative roles in gene transcription. Recently, we have shown that dMi-2 is recruited to the hsp70 gene in a heat shock-dependent manner, and is required to achieve high transcript levels. Here, we use chromatin immunoprecipitation sequencing (ChIP-Seq) to identify other chromatin regions displaying increased dMi-2 binding upon heat shock and to characterize the distribution of dMi-2 over heat shock genes. We show that dMi-2 is recruited to the body of at least seven heat shock genes. Interestingly, dMi-2 binding extends several hundred base pairs beyond the polyadenylation site into the region where transcriptional termination occurs. We find that dMi-2 does not associate with the entire nucleosome-depleted hsp70 locus 87A. Rather, dMi-2 binding is restricted to transcribed regions. Our results suggest that dMi-2 distribution over active heat shock genes are determined by transcriptional activity.

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Redistribution of RNA polymerase II and dMi-2 on polytene chromosomes after heat shock. Immunofluorescence staining of polytene chromosomes with (A) RNA polymerase II (pol IIser2) antibody and DAPI and (B) dMi-2 antibody and DAPI. (Upper panels) no heat shock (NHS); (lower panels) heat shock (HS) conditions. Position of hsp70 loci 87A and 87C are indicated by arrows.
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gks178-F1: Redistribution of RNA polymerase II and dMi-2 on polytene chromosomes after heat shock. Immunofluorescence staining of polytene chromosomes with (A) RNA polymerase II (pol IIser2) antibody and DAPI and (B) dMi-2 antibody and DAPI. (Upper panels) no heat shock (NHS); (lower panels) heat shock (HS) conditions. Position of hsp70 loci 87A and 87C are indicated by arrows.

Mentions: During HS, RNAP II and elongation factors loose association with many actively transcribed genes and strongly accumulate at several HS loci (19–22). As a consequence, expression of many genes is down-regulated, whereas transcription of HS genes is strongly stimulated. This dramatic redistribution of RNAP II and elongation factors can be visualized by indirect immunofluorescence of polytene chromosomes. An antibody recognizing the elongating form of RNAP II (RNAP IIser2P) stains more than 100 transcriptionally active regions of polytene chromosomes at normal temperatures (Figure 1A, upper panel). In contrast, less than 10 strongly staining regions are revealed by the same antibody on polytene chromosomes after 20 min of HS (Figure 1A, lower panel). These regions include the hsp70 containing 87 A and 87C loci.Figure 1.


Recruitment of the ATP-dependent chromatin remodeler dMi-2 to the transcribed region of active heat shock genes.

Mathieu EL, Finkernagel F, Murawska M, Scharfe M, Jarek M, Brehm A - Nucleic Acids Res. (2012)

Redistribution of RNA polymerase II and dMi-2 on polytene chromosomes after heat shock. Immunofluorescence staining of polytene chromosomes with (A) RNA polymerase II (pol IIser2) antibody and DAPI and (B) dMi-2 antibody and DAPI. (Upper panels) no heat shock (NHS); (lower panels) heat shock (HS) conditions. Position of hsp70 loci 87A and 87C are indicated by arrows.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3367206&req=5

gks178-F1: Redistribution of RNA polymerase II and dMi-2 on polytene chromosomes after heat shock. Immunofluorescence staining of polytene chromosomes with (A) RNA polymerase II (pol IIser2) antibody and DAPI and (B) dMi-2 antibody and DAPI. (Upper panels) no heat shock (NHS); (lower panels) heat shock (HS) conditions. Position of hsp70 loci 87A and 87C are indicated by arrows.
Mentions: During HS, RNAP II and elongation factors loose association with many actively transcribed genes and strongly accumulate at several HS loci (19–22). As a consequence, expression of many genes is down-regulated, whereas transcription of HS genes is strongly stimulated. This dramatic redistribution of RNAP II and elongation factors can be visualized by indirect immunofluorescence of polytene chromosomes. An antibody recognizing the elongating form of RNAP II (RNAP IIser2P) stains more than 100 transcriptionally active regions of polytene chromosomes at normal temperatures (Figure 1A, upper panel). In contrast, less than 10 strongly staining regions are revealed by the same antibody on polytene chromosomes after 20 min of HS (Figure 1A, lower panel). These regions include the hsp70 containing 87 A and 87C loci.Figure 1.

Bottom Line: Interestingly, dMi-2 binding extends several hundred base pairs beyond the polyadenylation site into the region where transcriptional termination occurs.We find that dMi-2 does not associate with the entire nucleosome-depleted hsp70 locus 87A.Rather, dMi-2 binding is restricted to transcribed regions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Molecular Biology and Tumor Research, Philipps-University, Emil-Mannkopff-Strasse 2, 35037 Marburg, Germany.

ABSTRACT
The ATP-dependent chromatin remodeler dMi-2 can play both positive and negative roles in gene transcription. Recently, we have shown that dMi-2 is recruited to the hsp70 gene in a heat shock-dependent manner, and is required to achieve high transcript levels. Here, we use chromatin immunoprecipitation sequencing (ChIP-Seq) to identify other chromatin regions displaying increased dMi-2 binding upon heat shock and to characterize the distribution of dMi-2 over heat shock genes. We show that dMi-2 is recruited to the body of at least seven heat shock genes. Interestingly, dMi-2 binding extends several hundred base pairs beyond the polyadenylation site into the region where transcriptional termination occurs. We find that dMi-2 does not associate with the entire nucleosome-depleted hsp70 locus 87A. Rather, dMi-2 binding is restricted to transcribed regions. Our results suggest that dMi-2 distribution over active heat shock genes are determined by transcriptional activity.

Show MeSH
Related in: MedlinePlus