title: Drosophila histone locus bodies form by hierarchical recruitment of components.
abstract: Nuclear bodies are protein- and RNA-containing structures that participate in a wide range of processes critical to genome function. Molecular self-organization is thought to drive nuclear body formation, but whether this occurs stochastically or via an ordered, hierarchical process is not fully understood. We addressed this question using RNAi and proteomic approaches in Drosophila melanogaster to identify and characterize novel components of the histone locus body (HLB), a nuclear body involved in the expression of replication-dependent histone genes. We identified the transcription elongation factor suppressor of Ty 6 (Spt6) and a homologue of mammalian nuclear protein of the ataxia telangiectasia-mutated locus that is encoded by the homeotic gene multisex combs (mxc) as novel HLB components. By combining genetic manipulation in both cell culture and embryos with cytological observations of Mxc, Spt6, and the known HLB components, FLICE-associated huge protein, Mute, U7 small nuclear ribonucleoprotein, and MPM-2 phosphoepitope, we demonstrated sequential recruitment and hierarchical dependency for localization of factors to HLBs during development, suggesting that ordered assembly can play a role in nuclear body formation.
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Anne E
authors.initials: AE
authors.lastname: White
authors.forename: Brandon D
authors.initials: BD
authors.lastname: Burch
authors.forename: Xiao-Cui
authors.initials: XC
authors.lastname: Yang
authors.forename: Pamela Y
authors.initials: PY
authors.lastname: Gasdaska
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Dominski
authors.forename: William F
authors.initials: WF
authors.lastname: Marzluff
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Duronio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002460:Cell Line; D022003:Cell Nucleus Structures / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000378:metabolism; D005786:Gene Expression Regulation; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D010445:Peptide Elongation Factors / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D063146:Polycomb-Group Proteins; D040901:Proteomics / Q000379:methods; D034622:RNA Interference; D012322:RNA Precursors / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D020885:Ribonucleoprotein, U7 Small Nuclear / Q000378:metabolism; D014162:Transfection
chemical_list: D051017:Apoptosis Regulatory Proteins; D029721:Drosophila Proteins; D051858:Forkhead Transcription Factors; D006657:Histones; D010445:Peptide Elongation Factors; D010750:Phosphoproteins; D063146:Polycomb-Group Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D020885:Ribonucleoprotein, U7 Small Nuclear; C470423:Spt6 protein, Drosophila