title: Genome-Wide Analysis of Drosophila RBf2 Protein Highlights the Diversity of RB Family Targets and Possible Role in Regulation of Ribosome Biosynthesis.
abstract: RBf2 is a recently evolved retinoblastoma family member in Drosophila that differs from RBf1, especially in the C-terminus. To investigate whether the unique features of RBf2 contribute to diverse roles in gene regulation, we performed chromatin immunoprecipitation sequencing for both RBf2 and RBf1 in embryos. A previous model for RB-E2F interactions suggested that RBf1 binds dE2F1 or dE2F2, whereas RBf2 is restricted to binding to dE2F2; however, we found that RBf2 targets approximately twice as many genes as RBf1. Highly enriched among the RBf2 targets were ribosomal protein genes. We tested the functional significance of this finding by assessing RBf activity on ribosomal protein promoters and the endogenous genes. RBf1 and RBf2 significantly repressed expression of some ribosomal protein genes, although not all bound genes showed transcriptional effects. Interestingly, many ribosomal protein genes are similarly targeted in human cells, indicating that these interactions may be relevant for control of ribosome biosynthesis and growth. We carried out bioinformatic analysis to investigate the basis for differential targeting by these two proteins and found that RBf2-specific promoters have distinct sequence motifs, suggesting unique targeting mechanisms. Association of RBf2 with these promoters appears to be independent of dE2F2/dDP, although promoters bound by both RBf1 and RBf2 require dE2F2/dDP. The presence of unique RBf2 targets suggest that evolutionary appearance of this corepressor represents the acquisition of potentially novel roles in gene regulation for the RB family.
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319.
authors.forename: Yiliang
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Laboratory of Molecular Neurobiology and Laboratory of Bioinformatics, Nencki Institute of Experimental Biology, Pasteura 3, 02-093 Warsaw, Poland.
authors.forename: Shamba S
authors.initials: SS
authors.lastname: Mondal
authors.affiliation: Cell and Molecular Biology Program, Michigan State University, East Lansing, Michigan 48824-1319.
authors.forename: Rima
authors.initials: R
authors.lastname: Mouawad
authors.affiliation: Institute of Informatics, University of Warsaw, Banacha 2, 02-097 Warsaw, Poland.
authors.forename: Bartek
authors.initials: B
authors.lastname: Wilczyński
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319.
authors.forename: R William
authors.initials: RW
authors.lastname: Henry
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319 arnosti@msu.edu.
authors.forename: David N
authors.initials: DN
authors.lastname: Arnosti
mesh_terms: D000818:Animals; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004330:Drosophila / Q000235:genetics* / Q000254:growth & development; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D050703:E2F2 Transcription Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005136:Eye Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D016678:Genome; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D016160:Retinoblastoma Protein; D012270:Ribosomes / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: C452919:BEAF-32 protein, Drosophila; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D050703:E2F2 Transcription Factor; C496298:E2f1 protein, Drosophila; C115434:E2f2 protein, Drosophila; D005136:Eye Proteins; C467582:RBF2 protein, Drosophila; D012330:RNA, Double-Stranded; C099965:Rbf protein, Drosophila; D012097:Repressor Proteins; D016160:Retinoblastoma Protein; D014157:Transcription Factors
keywords: Drosophila; RBf1; RBf2; retinoblastoma; ribosomal protein gene