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J Biol Chem, Vol. 273, Issue 13, 7268-7276, March 27, 1998
From the There is currently no published report on the
isolation and definitive identification of histone H1 in
Saccharomyces cerevisiae. It was, however, recently shown
that the yeast HHO1 gene codes for a predicted protein
homologous to H1 of higher eukaryotes (Landsman, D. (1996) Trends
Biochem. Sci. 21, 287-288; Ushinsky, S. C., Bussey, H.,
Ahmed, A. A., Wang, Y., Friesen, J., Williams, B. A., and
Storms, R. K. (1997) Yeast 13, 151-161), although
there is no biochemical evidence that shows that Hho1p is, indeed,
yeast histone H1. We showed that purified recombinant Hho1p (rHho1p) has electrophoretic and chromatographic properties similar to linker
histones. The protein forms a stable ternary complex with a
reconstituted core di-nucleosome in vitro at molar
rHho1p:core ratios up to 1. Reconstitution of rHho1p with H1-stripped
chromatin confers a kinetic pause at ~168 base pairs in the
micrococcal nuclease digestion pattern of the chromatin. These results
strongly suggest that Hho1p is a bona fide linker histone. We deleted
the HHO1 gene and showed that the strain is viable and has
no growth or mating defects. Hho1p is not required for telomeric
silencing, basal transcriptional repression, or efficient sporulation.
Unlike core histone mutations, a hho1
The Biochemical and Phenotypic Characterization of Hho1p, the
Putative Linker Histone H1 of Saccharomyces cerevisiae
,
Department of Biochemistry and Molecular
Biology, Pennsylvania State University,
University Park, Pennsylvania 16802, ¶ University of
Massachusetts Medical Center, Program in Molecular Medicine and
Department of Biochemistry and Molecular Biology,
Worcester, Massachusetts 01605, and ** National Center for
Biotechnology Information, National Library of Medicine, National
Institutes of Health, Bethesda, Maryland 20894
strain does not
exhibit a Sin or Spt phenotype. The absence of Hho1p does not lead to a
change in the nucleosome repeat length of bulk chromatin nor to
differences in the in vivo micrococcal nuclease cleavage
sites in individual genes as detected by primer extension mapping.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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