The drosophila zinc finger Protein CG9609 interacts with the Deubiquitinating (DUB) module of the SAGA complex and participates in the regulation of Transcription

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Abstract

In previous studies, we found that the zinc finger proteins Su(Hw) and CG9890 interact with the Drosophila SAGA complex and participate in the formation of active chromatin structure and transcriptional regulation. In this research, we discovered the interaction of the DUB module of the SAGA complex with another zinc finger protein, CG9609. ChIP-Seq analysis was performed and CG9609 binding sites in the Drosophila genome were identified. Analysis of binding sites showed that they are localized predominantly at gene promoters. The CG9609 protein has been shown to be involved in the regulation of gene expression.

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About the authors

J. V. Nikolenko

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: krasnov@genebiology.ru
Russian Federation, Moscow, 119991

M. M. Kurshakova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: krasnov@genebiology.ru
Moscow, 119991

D. V. Kopytova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: krasnov@genebiology.ru
Russian Federation, Moscow, 119991

Y. A. Vdovina

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: krasnov@genebiology.ru
Russian Federation, Moscow, 119991

N. E. Vorobyova

Institute of Gene Biology, Russian Academy of Sciences

Email: krasnov@genebiology.ru
Russian Federation, Moscow, 119334

A. N. Krasnov

Institute of Gene Biology, Russian Academy of Sciences

Author for correspondence.
Email: krasnov@genebiology.ru
Russian Federation, Moscow, 119334

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Supplementary files

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3. Fig. 1. Coimmunoprecipitation of 3xFLAG_CG9609 protein with ENY2 and Sgf11 proteins. The antibodies used in immunoprecipitation are indicated. Input is the initial extract, IgG is immunoprecipitation with nonspecific antibodies. Western blot was stained with antibodies to the 3xFLAG epitope. The 3xFLAG_CG9609 protein is marked with an arrow. It is seen that antibodies to ENY2 and Sgf11 precipitate 3xFLAG_CG9609. In the IgG sample, the wide band corresponds to the heavy chain of the antibodies; the 3xFLAG_CG9609 protein is absent.

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4. Fig. 2. ChIP-Seq profile of CG9609 protein in the bnl gene region. The upper panel shows the gene structure from the genome browser. The lower panel shows the ChIP-Seq profile itself in CPM units. It is evident that CG9609 protein is localized on the bnl gene promoter.

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5. Fig. 3. Consensus sequence of the potential binding site of the CG9609 protein.

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6. Fig. 4. Transcription of some genes containing CG9609 on promoters during RNA interference of CG9609. Gene names are given at the bottom. Light columns correspond to gene transcription under normal conditions (taken as one). Dark columns correspond to gene transcription during RNA interference of CG9609. The ordinate axis shows the ratio of transcription in the experiment and in the control. The level of CG9609 gene transcription during RNA interference was 9 times lower than under normal conditions. The ras64B gene was used as a normalization gene. The experiments were performed in triplicate. Error bars indicate the standard error of the mean. Statistically significant changes are indicated by an asterisk (p < 0.05, Student's t-test).

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