Role for a Drosophila Myb-containing protein complex in site-specific DNA replication

Eileen L Beall; J Robert Manak; Sharleen Zhou; Maren Bell; Joseph S Lipsick; Michael R Botchan

doi:10.1038/nature01228

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Role for a Drosophila Myb-containing protein complex in site-specific DNA replication

Journal article

Peer reviewed

Role for a Drosophila Myb-containing protein complex in site-specific DNA replication

Eileen L Beall, J Robert Manak, Sharleen Zhou, Maren Bell, Joseph S Lipsick and Michael R Botchan

Nature (London), Vol.420(6917), pp.833-837

12/19/2002

DOI: 10.1038/nature01228

PMID: 12490953

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Abstract

There is considerable interest in the developmental, temporal and tissue-specific patterns of DNA replication in metazoans. Site-specific DNA replication at the chorion loci in Drosophila follicle cells leads to extensive gene amplification, and the organization of the cis-acting DNA elements that regulate this process may provide a model for how such regulation is achieved. Two elements important for amplification of the third chromosome chorion gene cluster, ACE3 and Ori-beta, are directly bound by Orc (origin recognition complex), and two-dimensional gel analysis has revealed that the primary origin used is Ori-beta (refs 7-9). Here we show that the Drosophila homologue of the Myb (Myeloblastosis) oncoprotein family is tightly associated with four additional proteins, and that the complex binds site-specifically to these regulatory DNA elements. Drosophila Myb is required in trans for gene amplification, showing that a Myb protein is directly involved in DNA replication. A Drosophila Myb binding site, as well as the binding site for another Myb complex member (p120), is necessary in cis for replication of reporter transgenes. Chromatin immunoprecipitation experiments localize both proteins to the chorion loci in vivo. These data provide evidence that specific protein complexes bound to replication enhancer elements work together with the general replication machinery for site-specific origin utilization during replication.

DNA Replication

Chromatin - metabolism

Origin Recognition Complex

Genes, Insect - genetics

Transgenes - genetics

Substrate Specificity

DNA-Binding Proteins - genetics

Drosophila Proteins - metabolism

Drosophila melanogaster - genetics

Precipitin Tests

Proto-Oncogene Proteins c-myb - metabolism

Regulatory Sequences, Nucleic Acid - genetics

Macromolecular Substances

DNA Footprinting

DNA-Binding Proteins - metabolism

Drosophila melanogaster - metabolism

Animals

Gene Amplification

Replication Origin

Protein Binding

Female

Enhancer Elements, Genetic - genetics

Binding Sites

Chromatin - genetics

Details

Title: Subtitle: Role for a Drosophila Myb-containing protein complex in site-specific DNA replication
Creators: Eileen L Beall - Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA
J Robert Manak
Sharleen Zhou
Maren Bell
Joseph S Lipsick
Michael R Botchan
Resource Type: Journal article
Publication Details: Nature (London), Vol.420(6917), pp.833-837
DOI: 10.1038/nature01228
PMID: 12490953
NLM abbreviation: Nature
ISSN: 0028-0836
eISSN: 1476-4687
Publisher: England
Language: English
Date published: 12/19/2002
Academic Unit: Stead Family Department of Pediatrics; Biology; Craniofacial Anomalies Research Center
Record Identifier: 9983991991202771

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