Fractionation of human H1 subtypes and characterization of a subtype-specific antibody exhibiting non-uniform nuclear staining

Parseghian, Missag H.; Clark, Robert F.; Hauser, Loren; Dvorkin, Nadja; Harris, Debra A.; Hamkalo, Barbara A.

doi:10.1007/bf00710036

Cited by 36 publications

(28 citation statements)

References 72 publications

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“…We first digested the mixture of purified calf thymus linker histones with chymotrypsin, which cleaves preferentially at a unique conserved phenylalanine (Phe-108 in H1.1) in the globular domain near the C-tail (see Fig. 2A, asterisks) (63). Next, aliquots were removed from the chymotrypsin reaction at different times and quenched by adding SDS-PAGE loading buffer.…”

Section: Resultsmentioning

confidence: 99%

Binding of Barrier to Autointegration Factor (BAF) to Histone H3 and Selected Linker Histones Including H1.1

Lee

Wilson

2005

Journal of Biological Chemistry

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Barrier to autointegration factor (BAF) is an essential conserved double-stranded DNA-binding protein in metazoans. BAF binds directly to LEM domain nuclear proteins (e.g. LAP2, Emerin, and MAN1), lamin A, homeodomain transcription factors, and human immunodeficiency virus type 1-encoded proteins. BAF influences higher order chromatin structure and is required to assemble nuclei. BAF also facilitates retroviral preintegration complex insertion into target DNA in vitro, through unknown mechanisms. We report that BAF binds directly and selectively to linker histone H1.1 (among three subtypes tested) and core histone H3 with affinities of ϳ700 nM and ϳ100 -200 nM, respectively, in vitro and in vivo. Mutations at the bottom and top surfaces of the BAF dimer disrupted or enhanced, respectively, this binding and affected H1 and H3 similarly. Biochemical studies showed that C-terminal residues 108 -215 of histone H1.1 and the N-terminal tail plus helix ␣N in the core of histone H3.1 were each necessary and sufficient to bind BAF. Based on its interactions with histones and DNA, we propose BAF might bind nucleosomes in vivo.

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Section: Resultsmentioning

confidence: 99%

Binding of Barrier to Autointegration Factor (BAF) to Histone H3 and Selected Linker Histones Including H1.1

Lee

Wilson

2005

Journal of Biological Chemistry

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“…B Input samples (25 µg of HindIII/EcoRI-digested DNA-protein adducts) were treated with anti-RAP1 (lanes 1), anti-RNA polymerase II (lanes 2), a mixture of the three affinity-purified anti-linker histone IgGs (lanes 3) and probed with the sequences indicated below the panels gests that each of the common somatic H1s in mammals has distinct functions as has been argued on the basis of their evolutionary history (Ponte et al 1998). However, Parseghian et al (1993Parseghian et al ( , 1994 were unable to obtain an antibody against histone H1a and therefore were unable to localize all the human H1 variants. The localization pattern that we find in Arabidopsis is most similar to that of the midge (C. thummi) histones H1-II and -III, rat H1A and H1-B and human H1-1.…”

Section: Discussionmentioning

confidence: 96%

“…A similar set of results have been found in human cells, with H1A and H1-B binding throughout nuclei and H1°being restricted, being most abundant in nucleoli. The specific binding of the common H1 variants of human was investigated by Parseghian et al (1993Parseghian et al ( , 1994. They found that H1-1 (H1d) is uniformly distributed throughout nuclei, H1-2 and H1-4 (c and b) stain in a punctate pattern, and H1-3 (H1a) preferentially labels the nuclear periphery.…”

Section: Discussionmentioning

confidence: 99%

Subnuclear distribution of the entire complement of linker histone variants in Arabidopsis thaliana

Ascenzi

Gantt

1999

Chromosoma

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Linker histones (e.g. H1, H5, H1 degrees ) are thought to exert control on chromatin function by restricting nucleosomal dynamics. All higher eukaryotes possess a diverse family of linker histones, which may exhibit functional specialization. Arabidopsis thaliana apparently contains a minimal complement of linker histone structural variants and therefore is an ideal model for investigating functional differentiation among linker histones. Histones H1-1 and H1-2 are relatively similar proteins that are expressed in a wide variety of tissues and make up the majority of linker histone while H1-3 is a highly divergent minor variant protein that is induced by drought stress. We are interested in determining whether the in vivo distribution of each of these proteins also differs. To this end, we have produced subtype-specific antibodies and have localized each of the three proteins at the intranuclear and DNA sequence levels by indirect immunofluorescence and immunoprecipitation, respectively. Antibodies against linker histones H1-1 and H1-2 decorate nuclei in patterns very similar to 4',6-diamidino-2-phenylindole (DAPI) staining, but different than the staining pattern of total histones. In contrast, antibodies made against two regions of H1-3 bind to chromatin in a diffuse pattern distinct from the DAPI-staining pattern. We also describe a technique to determine the localization of plant linker histone variants along regions of chromatin, employing in vivo chemical DNA-protein cross-linking to preserve native associations followed by immunoprecipitation with subtype-specific antibodies. We use this technique to demonstrate that, in contrast to the major linker histones, H1-3 does not bind the repetitive sequences pAL1 and 5S rDNA. In addition, we show that linker histones are bound to the compacted nucleosomal arrays at the telomere but with reduced stoichiometry. Taken together, our results suggest that plants, as has been shown for animals, possess a variant linker histone that is differentially localized.

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“…Antibodies were purified using protein A-Sepharose (Pharmacia) and, if needed, subsequently affinity purified on filter strips containing the various bacterial fusion proteins. MeCP2 antibodies (22) and histone H1 antibodies (23) were kind gifts of Drs. A. Bird and M. Parseghian, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Given the presence in ZFP-37 of a histone H1-like domain, staining of ZFP-37 in neurons was compared with that of histone H1, using antibodies that recognize all common H1 isoforms (23). Like ZFP-37, histone H1 was found to have a variable intracellular localization (data not shown).…”

Section: Zfp-37 May Define Neuronal Nuclear Domains 9106mentioning

confidence: 99%

The Centromeric/Nucleolar Chromatin Protein ZFP-37 May Function to Specify Neuronal Nuclear Domains

Payen¹,

Verkerk²,

Michalovich³

et al. 1998

Journal of Biological Chemistry

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Murine ZFP-37 is a member of the large family of C 2 H 2 type zinc finger proteins. It is characterized by a truncated NH 2 -terminal Krü ppel-associated box and is thought to play a role in transcriptional regulation. During development Zfp-37 mRNA is most abundant in the developing central nervous system, and in the adult mouse expression is restricted largely to testis and brain. Here we show that at the protein level ZFP-37 is detected readily in neurons of the adult central nervous system but hardly in testis. In brain ZFP-37 is associated with nucleoli and appears to contact heterochromatin. Mouse and human ZFP-37 have a basic histone H1-like linker domain, located between KRAB and zinc finger regions, which binds double-stranded DNA. Thus we suggest that ZFP-37 is a structural protein of the neuronal nucleus which plays a role in the maintenance of specialized chromatin domains.

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Fractionation of human H1 subtypes and characterization of a subtype-specific antibody exhibiting non-uniform nuclear staining

Cited by 36 publications

References 72 publications

Binding of Barrier to Autointegration Factor (BAF) to Histone H3 and Selected Linker Histones Including H1.1

Binding of Barrier to Autointegration Factor (BAF) to Histone H3 and Selected Linker Histones Including H1.1

Subnuclear distribution of the entire complement of linker histone variants in Arabidopsis thaliana

The Centromeric/Nucleolar Chromatin Protein ZFP-37 May Function to Specify Neuronal Nuclear Domains

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