Developmental and cell cycle regulation of the Drosophila histone locus body.
Cyclin E/Cdk2 is necessary for replication-dependent histone mRNA biosynthesis, but how it controls this process in early development is unknown. We show that in Drosophila embryos the MPM-2 monoclonal antibody, raised against a phosphoepitope from human mitotic cells, detects Cyclin E/Cdk2-dependent nuclear foci that colocalize with nascent histone transcripts. These foci are coincident with the histone locus body (HLB), a Cajal body-like nuclear structure associated with the histone locus and enriched in histone pre-mRNA processing factors such as Lsm11, a core component of the U7 small nuclear ribonucleoprotein. Using MPM-2 and anti-Lsm11 antibodies, we demonstrate that the HLB is absent in the early embryo and occurs when zygotic histone transcription begins during nuclear cycle 11. Whereas the HLB is found in all cells after its formation, MPM-2 labels the HLB only in cells with active Cyclin E/Cdk2. MPM-2 and Lsm11 foci are present in embryos lacking the histone locus, and MPM-2 foci are present in U7 mutants, which cannot correctly process histone pre-mRNA. These data indicate that MPM-2 recognizes a Cdk2-regulated protein that assembles into the HLB independently of histone mRNA biosynthesis. HLB foci are present in histone deletion embryos, although the MPM-2 foci are smaller, and some Lsm11 foci are not associated with MPM-2 foci, suggesting that the histone locus is important for HLB integrity.
White
Anne E
AE
Department of Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
Leslie
Michelle E
ME
Calvi
Brian R
BR
Marzluff
William F
WF
Duronio
Robert J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002453:Cell Cycle; D022003:Cell Nucleus Structures / Q000378:metabolism*; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D004261:DNA Replication; D004331:Drosophila melanogaster / Q000166:cytology* / Q000196:embryology* / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D006657:Histones / Q000235:genetics*; D006801:Humans; D008938:Mitosis; D012322:RNA Precursors / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020885:Ribonucleoprotein, U7 Small Nuclear / Q000378:metabolism; D014158:Transcription, Genetic; D015053:Zygote / Q000378:metabolism
D000911:Antibodies, Monoclonal; D019927:Cyclin E; D006657:Histones; D012322:RNA Precursors; D012333:RNA, Messenger; D020885:Ribonucleoprotein, U7 Small Nuclear; D051357:Cyclin-Dependent Kinase 2