Early-late genes of the ecdysone cascade as models for transcriptional studies.
The DHR3 and Hr4 early-late genes of the ecdysone cascade are described as models for transcriptional studies in Drosophila cells. In a set of experiments, it became clear that these genes are a convenient and versatile system for research into the physiological conditions upon 20-hydroxyecdysone induction. DHR3 and Hr4 gene transcription is characterized by fast activation kinetics, which enables transcriptional studies without the influence of indirect effects. A limited number of activated genes (only 73 genes are induced one hour after treatment) promote the selectivity of transcriptional studies via 20-hydroxyecdysone induction. DHR3 and Hr4 gene expression is dose dependent, is completely controlled by the hormone titer and decreases within hours of 20-hydroxyecdysone withdrawal. The DHR3 and Hr4 gene promoters become functional within 20 minutes after induction, which makes them useful tools for investigation if the early activation process. Their transcription is controlled by the RNA polymerase II pausing mechanism, which is widespread in the genome of Drosophila melanogaster but is still underinvestigated. Uniform expression activation of the DHR3 and Hr4 genes in a cell population was confirmed at both the RNA and protein levels. Homogeneity of the transcription response makes DHR3/Hr4 system valuable for investigation of the protein dynamics during transcription induction.
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Marina Yu
MY
Mazina
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Julia V
JV
Nikolenko
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Nadezda A
NA
Fursova
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Petr N
PN
Nedil'ko
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Aleksey N
AN
Krasnov
a Department of Transcription Regulation and Chromatin Dynamic ; Institute of Gene Biology , Russian Academy of Sciences ; Moscow , Russia.
Nadezhda E
NE
Vorobyeva
D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004441:Ecdysterone / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D059014:High-Throughput Nucleotide Sequencing; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic
D029721:Drosophila Proteins; C502084:HR4 protein, Drosophila; C106186:Hr3 protein, Drosophila; D018160:Receptors, Cytoplasmic and Nuclear; D004441:Ecdysterone; D012319:RNA Polymerase II
20-hydroxyecdysone; RNA polymerase II; coactivator; early-late genes; transcription