The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo

The Drosophila protein Hrb57A has sequence homology to mammalian heterogenous nuclear ribonucleoprotein (hnRNP) K proteins. Its in vivo distribution has been studied at high resolution by confocal laser scanning microscopy (CLSM) in embryos injected with fluorescently labeled monoclonal antibody. Injection of antibody into living embryos had no apparent deleterious effects on further development. Furthermore, the antibody-protein complex could be observed for more than 7 cell cycles in vivo, revealing a dynamic redistribution from the nucleus to cytoplasm at each mitosis from blastoderm until hatching. The evaluation of two- and three-dimensional CLSM data sets demonstrated important differences in the localization of the protein in the nuclei of living compared to fixed embryos. The Hrb57A protein was recruited to the 93D locus upon heat shock and thus serves as an in vivo probe for the activity of the gene in diploid cells of the embryo. Observations during heat shock revealed considerable mobility within interphase nuclei of this transcription site. Furthermore, the reinitiation as well as the down regulation of transcriptional loci in vivo during the recovery from heat shock could be followed by the rapid redistribution of the hnRNP K during stress recovery. These data are incompatible with a model of the interphase nucleus in which transcription complexes are associated with a rigid nuclear matrix.

[1]  Prof. Dr. José A. Campos-Ortega,et al.  The Embryonic Development of Drosophila melanogaster , 1997, Springer Berlin Heidelberg.

[2]  M.,et al.  hnRNP proteins and B23 are the major proteins of the internal nuclear matrix of HeLa S3 cells , 1996, Journal of cellular biochemistry.

[3]  J. Strouboulis,et al.  Functional compartmentalization of the nucleus. , 1996, Journal of cell science.

[4]  D. Arndt-Jovin,et al.  FISH in whole‐mount Drosophila embryos. RNA: activation of a transcriptional locus, DNA: gene architecture and expression , 1996 .

[5]  D. Agard,et al.  Perturbation of Nuclear Architecture by Long-Distance Chromosome Interactions , 1996, Cell.

[6]  David Levens,et al.  Heterogeneous nuclear ribonucleoprotein K is a transcription factor , 1996, Molecular and cellular biology.

[7]  J. Dubochet,et al.  High-pressure freezing for cryoelectron microscopy. , 1995, Trends in cell biology.

[8]  H. Leffers,et al.  Characterisation of two major cellular poly(rC)-binding human proteins, each containing three K-homologous (KH) domains. , 1995, European journal of biochemistry.

[9]  M. Pardue,et al.  Stability of tandem repeats in the Drosophila melanogaster Hsr-omega nuclear RNA. , 1995, Genetics.

[10]  P. Park,et al.  Organization of centromeric domains in hepatocyte nuclei: rearrangement associated with de novo activation of the vitellogenin gene family in Xenopus laevis. , 1995, Experimental cell research.

[11]  M. Pardue,et al.  The nucleus-limited Hsr-omega-n transcript is a polyadenylated RNA with a regulated intranuclear turnover , 1994, The Journal of cell biology.

[12]  D. Hernandez-Verdun,et al.  The chromosome periphery during mitosis , 1994, BioEssays : news and reviews in molecular, cellular and developmental biology.

[13]  D. Arndt-Jovin,et al.  In vivo observation of the puff-specific protein no-on transient A (NONA) in nuclei of Drosophila embryos. , 1993, Journal of cell science.

[14]  B. Steensel,et al.  Fluorescent labeling of nascent RNA reveals transcription by RNA polymerase II in domains scattered throughout the nucleus , 1993, The Journal of cell biology.

[15]  J. Swedlow,et al.  Chromosome structure inside the nucleus. , 1993, Current opinion in cell biology.

[16]  D. Sahlas,et al.  Distribution of snRNPs, splicing factor SC-35 and actin in interphase nuclei: immunocytochemical evidence for differential distribution during changes in functional states. , 1993, Journal of cell science.

[17]  G. Dreyfuss,et al.  Association of individual hnRNP proteins and snRNPs with nascent transcripts , 1993, The Journal of cell biology.

[18]  D. Arndt-Jovin,et al.  Consequences of topoisomerase II inhibition in early embryogenesis of Drosophila revealed by in vivo confocal laser scanning microscopy. , 1993, Journal of cell science.

[19]  J. Lawrence,et al.  Higher level organization of individual gene transcription and RNA splicing. , 1993, Science.

[20]  D. Agard,et al.  The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis , 1993, The Journal of cell biology.

[21]  P. Park,et al.  Spatial rearrangement and enhanced clustering of kinetochores in interphase nuclei of dorsal root ganglion neurons in vitro: association with nucleolar fusion. , 1992, Experimental cell research.

[22]  G S Stein,et al.  Sequence-specific DNA-binding proteins are components of a nuclear matrix-attachment site. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Adrian R. Krainer,et al.  Regulation of alternative pre-mRNA splicing by hnRNP A1 and splicing factor SF2 , 1992, Cell.

[24]  G. Dreyfuss,et al.  Characterization of the major hnRNP proteins from Drosophila melanogaster , 1992, The Journal of cell biology.

[25]  G. Dreyfuss,et al.  Characterization and primary structure of the poly(C)-binding heterogeneous nuclear ribonucleoprotein complex K protein , 1992, Molecular and cellular biology.

[26]  E. Mack,et al.  Drosophila snRNP associated protein P11 which specifically binds to heat shock puff 93D reveals strong homology with hnRNP core protein A1. , 1991, Nucleic acids research.

[27]  M. Pardue,et al.  Expression of heat-shock locus hsr-omega in nonstressed cells during development in Drosophila melanogaster. , 1991, Developmental biology.

[28]  D. Johnson,et al.  The Drosophila Hrb87F gene encodes a new member of the A and B hnRNP protein group , 1991, Nucleic Acids Res..

[29]  A. Beyer,et al.  The Drosophila Hrb98DE locus encodes four protein isoforms homologous to the A1 protein of mammalian heterogeneous nuclear ribonucleoprotein complexes , 1990, Molecular and cellular biology.

[30]  M. Pardue,et al.  Multiple inducers of the Drosophila heat shock locus 93D (hsr omega): inducer-specific patterns of the three transcripts , 1989, The Journal of cell biology.

[31]  M. Pardue,et al.  A Drosophila heat shock locus with a rapidly diverging sequence but a conserved structure. , 1986, The Journal of biological chemistry.

[32]  M. Pardue,et al.  Heat shock locus 93D of Drosophila melanogaster: a spliced RNA most strongly conserved in the intron sequence. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[33]  G. Blobel,et al.  Gene gating: a hypothesis. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[34]  H. Weintraub,et al.  Localization of DNAase I-sensitive sequences to specific regions of interphase nuclei , 1985, Cell.

[35]  J. Sedat,et al.  Proteins shifting from the cytoplasm into the nuclei during early embryogenesis of Drosophila melanogaster. , 1984, Developmental biology.

[36]  C. D. Lewis,et al.  Interphase Nuclear Matrix and Metaphase Scaffolding Structures , 1984, Journal of Cell Science.

[37]  P. Kloetzel,et al.  Heat‐shock puff 93 D from Drosophila melanogaster: accumulation of a RNP‐specific antigen associated with giant particles of possible storage function. , 1983, The EMBO journal.

[38]  J. Sedat,et al.  Localization of antigenic determinants in whole Drosophila embryos. , 1983, Developmental biology.

[39]  M. Frasch,et al.  Nonpackaging and packaging proteins of hnRNA in Drosophila melanogaster , 1983, Cell.

[40]  B. Alberts,et al.  Studies of nuclear and cytoplasmic behaviour during the five mitotic cycles that precede gastrulation in Drosophila embryogenesis. , 1983, Journal of cell science.

[41]  E. Bautz,et al.  Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster. , 1983, The EMBO journal.

[42]  C. Allis,et al.  Mass isolation of pole cells from Drosophila melanogaster. , 1977, Developmental biology.

[43]  M. Pardue,et al.  The effect of heat shock on RNA synthesis in Drosophila tissues , 1976, Cell.

[44]  B. van Steensel,et al.  Nuclear domains and the nuclear matrix. , 1995, International review of cytology.

[45]  B. Brinkley,et al.  Nuclear matrix proteins as structural and functional components of the mitotic apparatus. , 1995, International review of cytology.

[46]  M. Kallajoki,et al.  Gel electrophoretic analysis of nuclear matrix fractions isolated from different human cell lines , 1994, Electrophoresis.

[47]  L. Jong,et al.  UvA-DARE ( Digital Academic Repository ) RNA polymerase II transcription is concentrated outside replication domains throughout S-phase , 1994 .

[48]  U. De Boni.  The interphase nucleus as a dynamic structure. , 1994, International review of cytology.

[49]  C. Burd,et al.  hnRNP proteins and the biogenesis of mRNA. , 1993, Annual review of biochemistry.

[50]  C Cremer,et al.  Role of chromosome territories in the functional compartmentalization of the cell nucleus. , 1993, Cold Spring Harbor symposia on quantitative biology.

[51]  D. Spector,et al.  Dynamics of transcription and pre-mRNA splicing within the mammalian cell nucleus. , 1993, Cold Spring Harbor symposia on quantitative biology.

[52]  N. Stuurman,et al.  Nuclear frameworks: concepts and operational definitions. , 1992, Cell biology international reports.

[53]  L. Ségalat,et al.  Spatial distribution of the Sm antigen in Drosophila early embryos , 1992, Biology of the cell.

[54]  K. Weber,et al.  Immunofluorescence and immunocytochemical procedures with affinity purified antibodies: tubulin-containing structures. , 1982, Methods in cell biology.