Skip to main content
SpringerLink
Log in

Characterization of a set of antibodies specific for three human histone H1 subtypes

  • Original Articles
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

A series of human histone H1 subtype-specific antibodies are described that were generated for localization and functional studies. Since our previous attempts to produce such antibodies against intact subtypes met with limited success, resulting in one antibody against a subtype we have designated H1-3, the approach used in the work presented is based on the production of antibodies against synthetic peptides or peptide fragments encompassing the variant NH2-terminal region of cach protein. Subtype-specific antibodies were obtained against synthetic peptides derived from subtypes designated H1-1 and H1-2 and the NH2-terminal fragment from an N-bromosuccinimide digest of H1–4. Antibody specificities were documented in all cases by enzymelinked immunosorbent and protein immunoblot assays against the purified subtypes as well as immunoblots against whole cell and nuclear extracts. In addition, the in vivo distribution of each antibody was determined by indirect immunofluorescence. H1-1 appears to be distributed in parallel with DNA concentration, similar to the results with an antibody that recognizes all subtypes, However, H1–2 and H1–4 are non-uniformly distributed, exhibiting similar punctate staining patterns. The staining patterns described are different from the pattern desciribed for the distribution of H1–3, suggesting that several subtypes are concentrated in distinct regions of the nucleus and, therefore, may be associated with distinct regions of the genome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ajiro K, Shibata K, Nishikawa Y (1990) Subtype-specific cyclic AMP-dependent histone H1 phosphorylation at the differentiation of mouse neuroblastoma cells. J Biol Chem 265: 6494–6500

    Google Scholar 

  • Allan J, Hartman PG, Crane-Robinson C, Aviles FY (1980) The structure of histone H1 and its location in chromatin. Nature 288: 675–679

    Google Scholar 

  • Banchev T, Srebreva L, Zlatanova J, Tsanev R (1988) Immunofluorescent localization of histone H10 in the nuclei of proliferating and differentiating friend cells. Exp Cell Res 177: 1–8

    Google Scholar 

  • Bartley J, Challey R (1970) Further studies of a thymus nucleohistone-associated protease. J Biol Chem 245: 4286–4292

    Google Scholar 

  • Bodanszky M (1988) Peptide chemistry: A practical textbook. Springer-Verlag, New York

    Google Scholar 

  • Boulikas T (1988) At least 60 ADP-ribosylated variant histones are present in nuclei from dimethylsulfate-treated and untreated cells. EMBO J 7: 57–67

    Google Scholar 

  • Boulikas T (1990) Studies on protein poly(ADP-ribosylation) using high resolution gel electroporesis. J Biol Chem 265: 14627–14631

    Google Scholar 

  • Boulikas T (1991) Relation between carcinogenesis, chromatin structure and poly (ADP-ribosylation). Anticancer Res 11:489–528

    Google Scholar 

  • Boulikas T, Bastin B, Boulikas P, Dupuis G (1990) Increase in histone poly(ADP-ribosylation) in mitogen-activated lymphoid cells. Exp Cell Res 187: 77–84

    Google Scholar 

  • Breneman JW, Yau P, Teplitz RL, Bradbury EM (1993) A light microscope study of linker histone distribution in rat metaphase chromosomes and interphase nuclei. Exp Cell Res 206: 16–26

    Google Scholar 

  • Campbell DH, Garvey JS, Cremer NE, Sussdorf DH (1970) Methods in immunology: A laboratory text for instruction and research, 2nd edn. Benjamin, New York

    Google Scholar 

  • Cohen LH, Newrock KM, Zweidler A (1975) Stage-specific switches in histone synthesis during embryogenesis of the sea urchin. Science 190: 994–997

    Google Scholar 

  • Cole RD (1987) Microheterogenity in H1 histones and its consequences. int J Pept Protein Res 30: 433–449

    Google Scholar 

  • Cole RD (1989) Purification and analysis of H1 histones. Methods Enzymol 170: 524–532

    Google Scholar 

  • D'Incalci M, Paola A, Wu RS, Bonner WM (1986) H1 variant synthesis in proliferating and quiescent human cells. Eur J Biochem 154: 273–279

    Google Scholar 

  • Dedon PC, Soults JA, Allis CD, Gorovsky MA (1991) Formaldehyde cross-linking and immunopecipitation demonstrate developmental changes in H1 association with transcriptionally active genes. Mol Cell Biol 11: 1729–1733

    Google Scholar 

  • Dignam JD, Lebovitz RM, Roeder RG (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11: 1475–1489

    Google Scholar 

  • Doenecke D, Toenjes R (1986) Differential distribution of lysine and arginine residues in the closely related histones H10 and H5. J Mol Biol 187: 461–464

    Google Scholar 

  • Goldman MA, Holmquist GP, Gray MC, Caston LA, Nag A (1984) Replication timing of genes and middle repetitive sequences. Science 224: 686–692

    Google Scholar 

  • Goodwin GH, Walker JM, Johns EW (1978) High mobility group nonhistone proteins. In: Busch H (ed) The cell nucleus: Chromatin, part C, vol VI. Academic Press, New York, pp 184–185

    Google Scholar 

  • Gorka C, Fakan S, Lawrence JJ (1993) Light and electron microscope immunocytochemical analyses of histone H10 distribution in the nucleus of friend erythroleukemia cells. Exp Cell Res 205: 152–158

    Google Scholar 

  • Harlow E, Lane D (1988) Antibodies: A laboratory manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY

    Google Scholar 

  • Higurashi M, Adachi H, Ohba Y (1987) Synthesis and degradation of H1 histone subtypes in mouse lymphoma L5178Y cells. J Biol Chem 262: 13075–13080

    Google Scholar 

  • Jackson DA, Hassan BA, Errington RJ, Cook PR (1993) Visualization of focal sites of transcription within human nuclei. EMBO J 12: 1059–1065

    Google Scholar 

  • Johnson DA, Gautsch JW, Sportman JR, Elder JH (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech 1: 3–8

    Google Scholar 

  • Kamakaka RT, Thomas JO (1990) Chromatin structure of transcriptionally competent and repressed genes. EMBO J 9: 3997–4006

    Google Scholar 

  • Kohn J, Wilchek M (1984) The use of cyanogen bromide and other novel cyanylating agents for the activation of polysaccharide resins. Appl Biochem Biotechnol 9: 285–304

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structureal proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • Lennox RW (1984) Differences in evolutionary stability among mammalian H1 subtypes: Implications for the roles of H1 subtypes om chromatin. J Biol Chem 259: 669–672

    Google Scholar 

  • Lennox RW, Cohen LH (1983) The histone H1 complements of dividing and nondividing cells of the mouse. J Biol Chem 258: 262–268

    Google Scholar 

  • Lennox RW, Oshima RG, Cohen LH (1982) The H1 histones and their interphase phosphorylated states in differentiated and undifferentiated cell lines derived from murine teratocarcinomas. J Biol Chem 257: 5183–5189

    Google Scholar 

  • Liao LW, Cole RD (1981 a) Condensation of dinucleosomes by individual subfractions of H1 histone. J Biol Chem 256: 10124–10128

    Google Scholar 

  • Liao LW, Cole RD (1981 b) Differnces among H1 histone subfractions in binding to linear and superhelical DNA: Sedimentation velocity studies. J Biol Chem 256: 11145–11150

    Google Scholar 

  • Mohr E, Trieschmann L, Grossbach U (1989) Histone H1 in two subspecies of Chironomus thummi with different genome sizes: Homologous chromosome sites differ largely in their content of a specific H1 variant. Proc Natl Acad Sci USA 86: 9308–9312

    Google Scholar 

  • Ohba Y, Higurashi M, Hayashi Y (1984) Phosphorylation of H1 subtypes in regenerating rat liver. J Biol Chem 259: 2942–2948

    Google Scholar 

  • Ohe Y, Hayashi H, Iwai K (1989) Human spleen histone H1. Isolation and amino acid sequences of three minor variants, H1a, H1c, and H1d. J Biochem (Tokyo) 106: 844–857

    Google Scholar 

  • Parseghian MH, Clark RF, Hauser LJ, Dvorkin N, Harris DA, Hamkalo BA (1993) Fractionation of human H1 subtypes and characterization of a subtype-specific antibody exhibiting nonuniform nuclear staining. Chromosome Res 1: 127–139

    Google Scholar 

  • Pehrson JR, Cohen LH (1985) Distribution of histone H1α among cells of the sea urchin embryo. Dev Biol 111: 530–533

    Google Scholar 

  • Pehrson JR, Cole RD (1982) Histone H1 subfractions and H10 tornover at different rates in nondividing cells. Biochemistry 21: 456–460

    Google Scholar 

  • Ramachandran LK, Witkop B (1967) N-Bromosuccinimide cleavage of peptides. Methods Enzymol XI: 283–299

    Google Scholar 

  • Rasheed BKA, Whisenant EC, Ghai RD, Papaionnou VE, Bhatnagar YM (1989) Biochemical and immunocytochemical analysis of a histone H1 variant from the mouse testis. J Cell Sci 94: 61–71

    Google Scholar 

  • Schulze E, Trieschmann L, Schulze B, Schmidt ER, Pitzel S, Zechel K, Grossbach U (1993) Structural and functional differences between histone H1 sequence variants with differential intranuclear distribution. Proc Natl Acad Sci USA 90: 2481–2485

    Google Scholar 

  • Seyedin SM, Cole RD, Kistler WS (1981) Histones from mammalian testes: The widespread occurrence of H1t. Exp Cell Res 136: 399–405

    Google Scholar 

  • Sherod D, Johnson G, Chalkley R (1974) Studies on the heterogenity of lysine-rich histones in dividing cells. J Biol Chem 249: 3923–3931

    Google Scholar 

  • Smerdon MJ, Isenberg I (1976) Interactions between the subfractions of calf thumus H1 and nonhistone chromosomal proteins HMG1 and HMG2. Biochemistry 15: 4242–4247

    Google Scholar 

  • Smith RC, Dworkin-Rastl E, Dworkin MB (1988) Expression of a histone H1-like protein is restricted to early Xenopus development. Genes Dev 2: 1284–1295

    Google Scholar 

  • Srebreva L, Zlatanova JS (1986) Specificity studies on anti-histone H1 antibodies obtaind by different immunization methods. FEBS Lett 200: 123–127

    Google Scholar 

  • Stone PR, Lorimer III WS, Kdwell WR (1977) Properties of the complex between histone H1 and poly (ADP-ribose) synthesized in HeLa cell nuclei. Eur J Biochem 81: 9–18

    Google Scholar 

  • Vaitukaitus J, Robbins JB, Nieschlag E, Ross GT (1971) A method for producing specific antisera with small doses of immunogen. J Clin Endocrinol 33: 988–991

    Google Scholar 

  • Wang JYJ (1988) Antibodies for phosphotyrosine: analytical and preparative tool for tyrosyl-phosphorylated proteins. Anal Biochem 172: 1–7

    Google Scholar 

  • Weintraub H (1984) Histone-H1-dependent chromatin superstructures and the suppression of gene activity. Cell 38: 17–27

    Google Scholar 

  • Wells DE (1986) Compilation analysis of histones and histone genes. Nucleic Acids Res 14 [Suppl]: r119-r149

    Google Scholar 

  • Wells DE, McBride C (1989) A comprehensive compilation and alignment of histones and histone genes. Nucleic Acids Res 17 [Suppl]: r311-r346

    Google Scholar 

  • Wong M, Kanai Y, Miwa M, Bustin M, Smulson M (1983) Immunological evidence for the in vivo occurrence of a crosslinked complex of poly (ADP-ribosylated) histone H1. Proc Natl Acad Sci USA 80: 205–209

    Google Scholar 

  • Wray W, Boulikas T, Wray VP, Hancok R (1981) Silver staining of proteins in polyacylamide gels. Anal Biochem 118: 197–203

    Google Scholar 

  • Zlatanova JS (1990) Immunochemical approaches to the study of histone H1 and high mobility group chromatin proteins. Mol Cell Biochem 92: 1–22

    Google Scholar 

  • Zlatanova JS, Srebreva LN, Banchev TB, Tasheva BT, Tsanev RG (1990) Cytoplasmic pool of histone H1 in mammalian cells. J Cell Sci 96: 461–468

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of Califonia, 92717, Irvine, CA, USA

    Missag H. Parseghian, Debra A. Harris, Darron R. Rishwain & Barbara A. Hamkalo

Authors
  1. Missag H. Parseghian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Debra A. Harris
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Darron R. Rishwain
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Barbara A. Hamkalo
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Parseghian, M.H., Harris, D.A., Rishwain, D.R. et al. Characterization of a set of antibodies specific for three human histone H1 subtypes. Chromosoma 103, 198–208 (1994). https://doi.org/10.1007/BF00368013

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00368013

Keywords

Search

Navigation