title: Concentrating pre-mRNA processing factors in the histone locus body facilitates efficient histone mRNA biogenesis.
abstract: The histone locus body (HLB) assembles at replication-dependent histone genes and concentrates factors required for histone messenger RNA (mRNA) biosynthesis. FLASH (Flice-associated huge protein) and U7 small nuclear RNP (snRNP) are HLB components that participate in 3' processing of the nonpolyadenylated histone mRNAs by recruiting the endonuclease CPSF-73 to histone pre-mRNA. Using transgenes to complement a FLASH mutant, we show that distinct domains of FLASH involved in U7 snRNP binding, histone pre-mRNA cleavage, and HLB localization are all required for proper FLASH function in vivo. By genetically manipulating HLB composition using mutations in FLASH, mutations in the HLB assembly factor Mxc, or depletion of the variant histone H2aV, we find that failure to concentrate FLASH and/or U7 snRNP in the HLB impairs histone pre-mRNA processing. This failure results in accumulation of small amounts of polyadenylated histone mRNA and nascent read-through transcripts at the histone locus. Thus, the HLB concentrates FLASH and U7 snRNP, promoting efficient histone mRNA biosynthesis and coupling 3' end processing with transcription termination.
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Deirdre C
authors.initials: DC
authors.lastname: Tatomer
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Esteban
authors.initials: E
authors.lastname: Terzo
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Kaitlin P
authors.initials: KP
authors.lastname: Curry
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Harmony
authors.initials: H
authors.lastname: Salzler
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Ivan
authors.initials: I
authors.lastname: Sabath
authors.affiliation: Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Zapotoczny
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599 Integrative Program for Biological and Genome Sciences, University of North Carolina, Chapel Hill, NC 27599 Department of Genetics, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: McKay
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599 Integrative Program for Biological and Genome Sciences, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Dominski
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599 Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599 Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599 Integrative Program for Biological and Genome Sciences, University of North Carolina, Chapel Hill, NC 27599 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599 marzluff@med.unc.edu duronio@med.unc.edu.
authors.forename: William F
authors.initials: WF
authors.lastname: Marzluff
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599 Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC 27599 Integrative Program for Biological and Genome Sciences, University of North Carolina, Chapel Hill, NC 27599 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599 Department of Genetics, University of North Carolina, Chapel Hill, NC 27599 marzluff@med.unc.edu duronio@med.unc.edu.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Duronio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics*; D056426:Genetic Loci; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D008954:Models, Biological; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020885:Ribonucleoprotein, U7 Small Nuclear / Q000378:metabolism; D019076:Transgenes
chemical_list: D002352:Carrier Proteins; D029721:Drosophila Proteins; C559982:FLASH protein, Drosophila; D006657:Histones; D050505:Mutant Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D020885:Ribonucleoprotein, U7 Small Nuclear