Linker histone protection of chromatosomes reconstituted on 5S rDNA from Xenopus borealis:a reinvestigation.
The location of the linker histone (LH) on the nucleosome has been the subject of recent controversy. Although previous evidence had supported a location over the dyad axis, some recent experiments suggest an asymmetric, off-axis position. In this paper we show that the DNA sequence used to reconstitute chromatosomes in these experiments is prone to artifacts in nuclease digestion: results interpreted as 'protection' by LHs can be obtained with either naked DNA or with reconstituted core nucleosomes, in the absence of LHs. Consequently, we feel that general interpretation or extrapolation of such results must be regarded with the utmost caution. In addition, we show that the protection data on an alternative, previously unreported major core position on this same DNA sequence support a model of asymmetric, off-axis position of the LH, with linker DNA protection on only one side of the core particle.
An
W
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Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331-7305, USA.
van Holde
K
K
Zlatanova
J
J
D000818:Animals; D016477:Artifacts; D018983:DNA Footprinting; D004275:DNA, Ribosomal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008836:Micrococcal Nuclease / Q000378:metabolism; D008957:Models, Genetic; D009707:Nucleosomes / Q000378:metabolism*; D011485:Protein Binding; D012341:RNA, Ribosomal, 5S; D014981:Xenopus
D004275:DNA, Ribosomal; D004268:DNA-Binding Proteins; D006657:Histones; D009707:Nucleosomes; D012341:RNA, Ribosomal, 5S; D008836:Micrococcal Nuclease