TY - JOUR

T1 - Tethering of CHROMATOR and dCTCF proteins results in decompaction of condensed bands in the Drosophila melanogaster polytene chromosomes but does not affect their transcription and replication timing

AU - Pokholkova, Galina V.

AU - Demakov, Sergei A.

AU - Andreenkov, Oleg V.

AU - Andreenkova, Natalia G.

AU - Volkova, Elena I.

AU - Belyaeva, Elena S.

AU - Zhimulev, Igor F.

PY - 2018/4/1

Y1 - 2018/4/1

N2 - Instulator proteins are central to domain organization and gene regulation in the genome. We used ectopic tethering of CHROMATOR (CHRIZ/CHRO) and dCTCF to pre-defined regions of the genome to dissect the influence of these proteins on local chromatin organization, to analyze their interaction with other key chromatin proteins and to evaluate the effects on transcription and replication. Specifically, using UAS-GAL4DBD system, CHRO and dCTCF were artificially recruited into highly compacted polytene chromosome bands that share the features of silent chromatin type known as intercalary heterochromatin (IH). This led to local chromatin decondensation, formation of novel DHSes and recruitment of several "open chromatin" proteins. CHRO tethering resulted in the recruitment of CP190 and Z4 (PZG), whereas dCTCF tethering attracted CHRO, CP190, and Z4. Importantly, formation of a local stretch of open chromatin did not result in the reactivation of silent marker genes yellow and mini-white immediately adjacent to the targeting region (UAS), nor did RNA polII become recruited into this chromatin. The decompacted region retained late replicated, similarly to the wild-type untargeted region.

AB - Instulator proteins are central to domain organization and gene regulation in the genome. We used ectopic tethering of CHROMATOR (CHRIZ/CHRO) and dCTCF to pre-defined regions of the genome to dissect the influence of these proteins on local chromatin organization, to analyze their interaction with other key chromatin proteins and to evaluate the effects on transcription and replication. Specifically, using UAS-GAL4DBD system, CHRO and dCTCF were artificially recruited into highly compacted polytene chromosome bands that share the features of silent chromatin type known as intercalary heterochromatin (IH). This led to local chromatin decondensation, formation of novel DHSes and recruitment of several "open chromatin" proteins. CHRO tethering resulted in the recruitment of CP190 and Z4 (PZG), whereas dCTCF tethering attracted CHRO, CP190, and Z4. Importantly, formation of a local stretch of open chromatin did not result in the reactivation of silent marker genes yellow and mini-white immediately adjacent to the targeting region (UAS), nor did RNA polII become recruited into this chromatin. The decompacted region retained late replicated, similarly to the wild-type untargeted region.

KW - Animals

KW - Animals, Genetically Modified

KW - CCCTC-Binding Factor/genetics

KW - Cell Cycle Proteins/genetics

KW - DNA Replication Timing

KW - Deoxyribonuclease I/metabolism

KW - Drosophila Proteins/genetics

KW - Drosophila melanogaster/genetics

KW - Microtubule-Associated Proteins/genetics

KW - Nuclear Matrix-Associated Proteins/genetics

KW - Nuclear Proteins/genetics

KW - Polytene Chromosomes/genetics

KW - Transcription, Genetic

KW - CHROMODOMAIN PROTEIN

KW - DECONDENSATION

KW - GENOME

KW - DNA-REPLICATION

KW - INSULATOR PROTEINS

KW - ORGANIZATION

KW - GENE-EXPRESSION

KW - INTERCALARY HETEROCHROMATIN

KW - DOMAINS

KW - UNDERREPLICATION

UR - http://www.scopus.com/inward/record.url?scp=85044855588&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0192634

DO - 10.1371/journal.pone.0192634

M3 - Article

C2 - 29608600

AN - SCOPUS:85044855588

VL - 13

SP - e0192634

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 4

M1 - e0192634

ER -