Biochemistry (Moscow)

, Volume 77, Issue 11, pp 1285–1293

Mapping of cis-regulatory sites in the promoter of testis-specific Stellate genes of Drosophila melanogaster

  • O. M. Olenkina
  • K. S. Egorova
  • A. A. Aravin
  • N. M. Naumova
  • V. A. Gvozdev
  • L. V. Olenina
Article

Abstract

Tandem Stellate genes organized into two clusters in heterochromatin and euchromatin of the X-chromosome are part of the Ste-Su(Ste) genetic system required for maintenance of male fertility and reproduction of Drosophilamelanogaster. Stellate genes encode a regulatory subunit of protein kinase CK2 and are the main targets of germline-specific piRNA-silencing; their derepression leads to appearance of protein crystals in spermatocytes, meiotic disturbances, and male sterility. A short promoter region of 134 bp appears to be sufficient for testis-specific transcription of Stellate, and it contains three closely located cis-regulatory elements called E-boxes. By using reporter analysis, we confirmed a strong functionality of the E-boxes in the Stellate promoter for in vivo transcription. Using selective mutagenesis, we have shown that the presence of the central E-box 2 is preferable to maintain a high-level testis-specific transcription of the reporter gene under the Stellate promoter. The Stellate promoter provides transcription even in heterochromatin, and corresponding mRNAs are translated with the generation of full-size protein products in case of disturbances in the piRNA-silencing process. We have also shown for the first time that the activity of the Stellate promoter is determined by chromatin context of the X-chromosome in male germinal cells, and it increases at about twofold when relocating in autosomes.

Key words

promoter spermatogenesis Drosophila Stellate genes heterochromatin 

Abbreviations

CK2

protein kinase 2 (casein kinase 2)

DPE

downstream promoter element

HLH

helix-loop-helix protein family

MSL

male-specific lethal complex of Drosophila

ONPG

o-nitrophenyl-β-D-galactopyranoside

ORF

open reading frame

PCR

polymerase chain reaction

piRNAs

PIWI-interacting short regulatory RNAs

Su(Ste)

Suppressor of Stellate or crystal locus

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Copyright information

© Pleiades Publishing, Ltd. 2012

Authors and Affiliations

  • O. M. Olenkina
    • 1
  • K. S. Egorova
    • 1
  • A. A. Aravin
    • 1
  • N. M. Naumova
    • 1
  • V. A. Gvozdev
    • 1
  • L. V. Olenina
    • 1
  1. 1.Institute of Molecular GeneticsRussian Academy of SciencesMoscowRussia

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