Complete reconstitution of bypass and blocking functions in a minimal artificial insulator from complex.

TitleComplete reconstitution of bypass and blocking functions in a minimal artificial insulator from complex.
Publication TypeJournal Article
Year of Publication2019
AuthorsKyrchanova, O, Sabirov, M, Mogila, V, Kurbidaeva, A, Postika, N, Maksimenko, O, Schedl, P, Georgiev, P
JournalProc Natl Acad Sci U S A
Volume116
Issue27
Pagination13462-13467
Date Published2019 07 02
ISSN1091-6490
KeywordsAnimals, Drosophila, Drosophila Proteins, Gene Expression Regulation, Genes, Insect, Insulator Elements
Abstract

<p>Boundaries in the complex (BX-C) delimit autonomous regulatory domains that drive parasegment-specific expression of the genes , and The boundary is located between the and domains and has two key functions: blocking cross-talk between these domains and at the same time promoting communication (boundary bypass) between and the promoter. Using a replacement strategy, we found that multimerized binding sites for the architectural proteins Pita, Su(Hw), and dCTCF function as conventional insulators and block cross-talk between the and domains; however, they lack bypass activity, and is unable to regulate Here we show that an ∼200-bp sequence of dHS1 from the boundary rescues the bypass defects of these multimerized binding sites. The dHS1 sequence is bound in embryos by a large multiprotein complex, Late Boundary Complex (LBC), that contains the zinc finger proteins CLAMP and GAF. Using deletions and mutations in critical GAGAG motifs, we show that bypass activity correlates with the efficiency of recruitment of LBC components CLAMP and GAF to the artificial boundary. These results indicate that LBC orchestrates long-distance communication between the regulatory domain and the gene, while the Pita, Su(Hw), and dCTCF proteins function to block local cross-talk between the neighboring regulatory domains and .</p>

DOI10.1073/pnas.1907190116
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID31209019
PubMed Central IDPMC6613175
Grant ListR35 GM126975 / GM / NIGMS NIH HHS / United States