Nuclear protein LEDGF/p75 recognizes supercoiled DNA by a novel DNA-binding domain

Kimiko M. Tsutsui, Kuniaki Sano, Osamu Hosoya, Tadashi Miyamoto, Ken Tsutsui

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Lens epithelium-derived growth factor (LEDGF) or p75 is a co-activator of general transcription and also involved in insertion of human immunodeficiency virus type I (HIV-1) cDNA into host cell genome, which occurs preferentially to active transcription units. These phenomena may share an underlying molecular mechanism in common. We report here that LEDGF/p75 binds negatively supercoiled DNA selectively over unconstrained DNA. We identified a novel DNA-binding domain in the protein and termed it 'supercoiled DNA-recognition domain' (SRD). Recombinant protein fragments containing SRD showed a preferential binding to supercoiled DNA in vitro. SRD harbors a characteristic cluster of lysine and glutamic/aspartic acid residues. A polypeptide mimicking the cluster (K9E9K9) also showed this specificity, suggesting that the cluster is an essential element for the supercoil recognition. eGFP-tagged LEDGF/p75 expressed in the nucleus distributed partially in transcriptionally active regions that were identified by immunostaining of methylated histone H3 (H3K4me3) or incorporation of Br-UTP. This pattern of localization was observed with SRD alone but abolished if the protein lacked SRD. Thus, these results imply that LEDGF/p75 guides its binding partners, including HIV-1 integrase, to the active transcription site through recognition of negative supercoils generated around it.

Original languageEnglish
Pages (from-to)5067-5081
Number of pages15
JournalNucleic acids research.
Volume39
Issue number12
DOIs
Publication statusPublished - Jul 2011

Fingerprint

Superhelical DNA
Nuclear Proteins
DNA
Uridine Triphosphate
lens epithelium-derived growth factor
Recombinant Proteins
Histones
Lysine
HIV-1
Glutamic Acid
Catalytic Domain
Complementary DNA
HIV
Genome
Peptides

ASJC Scopus subject areas

  • Genetics

Cite this

Nuclear protein LEDGF/p75 recognizes supercoiled DNA by a novel DNA-binding domain. / Tsutsui, Kimiko M.; Sano, Kuniaki; Hosoya, Osamu; Miyamoto, Tadashi; Tsutsui, Ken.

In: Nucleic acids research., Vol. 39, No. 12, 07.2011, p. 5067-5081.

Research output: Contribution to journalArticle

Tsutsui, Kimiko M. ; Sano, Kuniaki ; Hosoya, Osamu ; Miyamoto, Tadashi ; Tsutsui, Ken. / Nuclear protein LEDGF/p75 recognizes supercoiled DNA by a novel DNA-binding domain. In: Nucleic acids research. 2011 ; Vol. 39, No. 12. pp. 5067-5081.
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