Identification of a highly conserved region critical for the functionality of the CP190 protein in Drosophila melanogaster

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Abstract

The CP190 protein binds to both housekeeping gene promoters and insulator/boundary elements and plays a critical role in their activity. The aim of this work was to study the effect of deletions in highly conserved regions of the CP190 protein on its functionality. It was shown that deletion of the sequence from 664 to 700 aa leads to a lethal phenotype. Thus, a new region was identified in CP190 that plays an important role in the interaction of the CP190 protein with as yet unidentified partner proteins, stabilization of protein complexes formed by CP190 and their recruitment to chromatin.

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About the authors

L. S. Melnikova

Institute of Gene Biology, Russian Academy of Sciences

Author for correspondence.
Email: lsm73@mail.ru
Russian Federation, Moscow

K. I. Balagurov

Institute of Gene Biology, Russian Academy of Sciences

Email: lsm73@mail.ru
Russian Federation, Moscow

P. G. Georgiev

Institute of Gene Biology, Russian Academy of Sciences

Email: lsm73@mail.ru

Academician of the RAS

Russian Federation, Moscow

A. K. Golovnin

Institute of Gene Biology, Russian Academy of Sciences

Email: lsm73@mail.ru
Russian Federation, Moscow

References

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Supplementary files

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2. Fig. 1. Schematic representation of the homology between CP190 proteins in different Drosophila species based on multiple sequence alignment. Drosophila species for which protein homology was assessed are indicated on the left. Homology levels for individual amino acid residues are shown as boxes ranked by height in the following order: 100%, 80%, 60%, 20%, and no homology. The “–” symbol denotes amino acid regions that are not part of the consensus sequence. Domain legend: BTB – BTB/POZ domain; NLS – nuclear localization signal; D – asparagine-rich region; M – centrosome-interacting regions; Znf – zinc finger domains; Q – glutamine-rich region; D/E – C-terminal domain rich in glutamine and asparagine. The identified highly conserved regions (HCR) and domains are indicated by segments below the corresponding amino acid sequences.

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3. Fig. 2. Derivatives of the CP190 protein used in the work. The full-length CP190 protein is shown schematically. The measurement scale (in aa) is shown above the scheme. Domain designations are as in Fig. 1. The names of the derivatives are indicated on the left, the amino acid residues that limit the derivative forms are designated by numbers. The size of the derivatives is indicated by segments, and the dotted lines indicate internal deletions. The results obtained in the analysis of the survival and fertility of flies in the corresponding transgenic lines are shown to the right of the schemes: “+” – the flies survived and were fertile; “–” – the flies did not survive; “+/–” – the viability and fertility of the flies were reduced.

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4. Fig. 3. Analysis of expression of CP190 derivatives in transgenic lines. Protein extract was isolated from flies of transgenic lines expressing different derivatives of CP190, the names of which are indicated above the corresponding lanes. The membrane was sequentially stained with anti-FLAG antibodies (α FLAG, upper panel) to determine the level of CP190 expression and with anti-tubulin antibodies (α Tub, lower panel) as a loading control. The y1w1118 line (negative control) was used to control the specificity of antibodies to the FLAG epitope. Molecular masses in kDa are indicated on the right.

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