Architectural protein pita cooperates with dCTCF in organization of functional boundaries in bithorax complex

Olga Kyrchanova, Nikolay Zolotarev, Vladic Mogila, Oksana Maksimenko, Paul Schedl, Pavel Georgiev

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Boundaries in the Bithorax complex (BX-C) of Drosophila delimit autonomous regulatory domains that drive parasegment-specific expression of homeotic genes. BX-C boundaries have two crucial functions: they must block crosstalk between adjacent regulatory domains and at the same time facilitate boundary bypass. The C2H2 zinc-finger protein Pita binds to several BX-C boundaries, including Fab-7 and Mcp. To study Pita functions, we have used a boundary replacement strategy by substituting modified DNAs for the Fab-7 boundary, which is located between the iab-6 and iab-7 regulatory domains. Multimerized Pita sites block iab-6↔iab-7 crosstalk but fail to support iab-6 regulation of Abd-B (bypass). In the case of Fab-7, we used a novel sensitized background to show that the two Pita-binding sites contribute to its boundary function. Although Mcp is from BX-C, it does not function appropriately when substituted for Fab-7: it blocks crosstalk but does not support bypass. Mutation of the Mcp Pita site disrupts blocking activity and also eliminates dCTCF binding. In contrast, mutation of the Mcp dCTCF site does not affect Pita binding, and this mutant boundary retains partial function.

Original languageEnglish (US)
Pages (from-to)2663-2672
Number of pages10
JournalDevelopment (Cambridge)
Volume144
Issue number14
DOIs
StatePublished - Jul 15 2017

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Developmental Biology

Keywords

  • Architectural proteins
  • Chromatin organization
  • Insulator
  • Protein-protein interactions
  • Zinc-finger transcription factors

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