A survey of H1<sup>0</sup>‐ and H5‐like protein structure and distribution in higher and lower eukaryotes

Smith, Bryan John; Harris, Martin; Sigournay, Catherine M.; Mayes, E. L. V.; Bustin, Michael

doi:10.1111/j.1432-1033.1984.tb07916.x

Cited by 76 publications

(25 citation statements)

References 42 publications

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“…Elsewhere we presented evidence that a yeast analogue of histone H 1 could be detected immunologically as a contaminant in some HMG preparations (12). Positive immunochemical identification of H1-like proteins in yeast has also been reported by others (6,8). As a continuation of this work (12), we attempted the identification in the yeast genome of nucleotide sequences homologous to the H 1 gene from sea urchin.…”

supporting

confidence: 61%

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Miloshev

Zlatanova

1994

J. Gen. Appl. Microbiol.

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supporting

confidence: 61%

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Miloshev

Zlatanova

1994

J. Gen. Appl. Microbiol.

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“…In the few nuclei still isolatable at 12 hr after H1 induction, chromatin was more highly condensed than in normal yeast nuclei. This hypercondensation may be achieved by the concerted action ofthe exogenous H1 and of an endogenous Hl-like protein (5)(6)(7). Yeast DNA appears rather compacted even in the interphase nucleus, with a compaction factor (the ratio of the length of total genomic DNA to the diameter of the nucleus) of roughly 2000 (26).…”

mentioning

confidence: 99%

Low levels of exogenous histone H1 in yeast cause cell death.

Miloshev

Venkov

Holde

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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To elucidate the hbnctin of lysine-rich histone, yeast cells, which are believed to lack this histone, were transformed with an expression vector carrying the sea urchin histone Hi gene under control of an inducible promoter. Expression of hill-length protein was tested by Immunobloffig and the intracellular distribution was monitored by immunoelectron microscopy. Even low amounts of exogenous H1 led to dramatic changes in ir lluar morphology and cell death. The cells that survived had lost either the plasmid or the ability to express the exogenous protein. Thus, even low amounts of canonical histone Hi are lethal to yeast cells.Histone H1 is known to bind to the linker DNA between nucleosomes, sealing off two turns of DNA around the histone octamer (1, 2). It is also involved in the formation and maintenance of higher-order chromatin structures and inhibits transcription by an as yet unknown mechanism (3, 4). One possible approach to elucidate the function of this histone would be to introduce exogenous H1 into appropriate living cells and follow its effects on the cellular function of interest. Yeast seems to be the obvious choice for such experiments, for it is believed not to contain an endogenous typical H1. Although immunological evidence for Hl-like proteins in yeast has been reported (5-7), such molecules might be quite different from the linker histones of higher eukaryotes. Moreover, the putative yeast H1 homologue is probably present in very low amounts and may be predominantly or exclusively localized in heterochromatic regions or at telomeres.Recently, we (8, 9) and others (10) Western Blotting. Total yeast proteins or 5% perchloric acid-extracted cellular proteins were fractionated electrophoretically in SDS-containing polyacrylamide gels (16) and transferred to nitrocellulose paper by semidry blotting in a model IMM-1 apparatus (W.E.P. Company, Seattle) as recommended by the manufacturer. Anti-mouse liver histone H1 antiserum (5) was presaturated with yeast lysate from the host strain by incubating nitrocellulose sheets (4 cm x 7 cm) with the lysate overnight at 40C and then with 10 ml ofanti-H1 antiserum for 3 hr at 370C. The filter with the adsorbed antibodies was removed and the remaining serum was used for Western blotting, performed according to routine procedures (5). Presaturation ofthe serum significantly reduced the background immunoreaction. Dilution of the anti-Hi antiserum was 1000-fold, and the enzyme substrate was o-phenylenediamine.Electron Microscopy. Cells were fixed for 5 min in 3.7% formaldehyde and then with 1% glutaraldehyde overnight at room temperature. Fixed cells were embedded in Lowacryl K4M (17) and ultrathin sections were examined in a JEOL JEM 100B electron microscope.Immune Microscopy. The immunostaining was performed directly on the microscopic grids at room temperature: ultrathin sections of Lowacryl-embedded fixed cells were incubated for 1 hr with presaturated serum (see above), which was diluted 400-fold in phosphate-buffered saline (PBS), washed in PBS,...

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“…4. Previous reports indicated that the N terminus of the H1 0 proteins is blocked, restricting the determination of primary structure in that region (15,31). However, as shown in Table I, we did sequence all RP-HPLC fractions including the N-terminal fraction I of Fig.…”

Section: Fig 2 Au-page Of the Four Histone H1mentioning

confidence: 98%

The Microheterogeneity of the Mammalian H10Histone

Lindner

Sarg

Hoertnagl

et al. 1998

Journal of Biological Chemistry

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The linker histones H1, H5, and H1 0 are associated with the core histone-DNA complex and with the linker DNA between adjacent nucleosomes and are thought to modulate the condensation/decondensation of the chromatin fiber, thus influencing many nuclear activities such as transcription, replication, recombination, and DNA repair (2). H1 0 was first described in 1969 by Panyim and Chalkley (3, 4) as an H1-like protein present in mammalian tissues with little or no cellular proliferation and was later shown to increase at a terminal stage of differentiation (5-10). Some cells, however, accumulate significant amounts of the protein while still actively proliferating (11, 12) or accumulate it upon proliferation arrest without concomitant differentiation (12). In addition, H1 0 seems to be the only histone undergoing changes during malignant transformation (13). Recently, it was found that transformation of NIH 3T3 fibroblasts by c-Ha-ras Val12 oncogene causes chromatin decondensation accompanied by alterations in the content of histone H1 0 (14). All these findings suggest a role for H1 0 in the regulation of either cell proliferation or cellular differentiation.In every tissue in which H1 0 has been detected, two subfractions were present (15-18). It appears that these two H1 0 proteins, up to now named H1 0 a and H1 0 b, have specific individual functions in chromatin (15). The relative proportions of the two H1 0 forms seem to differ from tissue to tissue (15) and exhibit age-dependent changes in rat brain cortical neurons (17). The two H1 0 s are resolvable by ion-exchange chromatography on Bio-Rex 70 (16, 19) or acetic acid-urea gel electrophoresis (15-18). Most recently, Lindner et al. (20) developed a high performance capillary electrophoresis method allowing separation of H1 0 and its subfractions from other histone H1 subtypes. The two H1 0 proteins run coincidentally on sodium dodecyl sulfate-polyacrylamide gels, suggesting that the difference between them is one of charge and not of size (15). Since neither treatment with alkaline phosphatase nor exposure to alkaline conditions changed the separation of the H1 0 peak into two subfractions, phosphorylation and ADP-ribosylation have been ruled out as possible post-translational modifications responsible for the different forms (16,20,21). Although some investigators speculated that the two forms of H1 0 might be coded by different genes (15, 17), Doenecke et al. (22) found that the mammalian genomes contain only one H1 0 gene.To gain insight into the nature of the two H1 0 subfractions, we took advantage of a combined reversed phase high performance liquid chromatography (RP-HPLC) 1 /hydrophilic interaction liquid chromatography (HILIC) technique recently developed in our laboratory for separating acetylated core and phosphorylated H1 histones (23,24). By applying this two-step HPLC method human placenta histone H1 0 was resolved into four components, which were treated with cyanogen bromide and chymotrypsin. HILIC analysis of the peptide fragments

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A survey of H1⁰‐ and H5‐like protein structure and distribution in higher and lower eukaryotes

Cited by 76 publications

References 42 publications

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Low levels of exogenous histone H1 in yeast cause cell death.

The Microheterogeneity of the Mammalian H10Histone

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A survey of H10‐ and H5‐like protein structure and distribution in higher and lower eukaryotes

Cited by 76 publications

References 42 publications

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Existence of yeast genomic fragments hybridizing with the sea urchin histone H1 gene.

Low levels of exogenous histone H1 in yeast cause cell death.

The Microheterogeneity of the Mammalian H10Histone

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A survey of H1⁰‐ and H5‐like protein structure and distribution in higher and lower eukaryotes