Histone redistribution and conformational effect on chromatin induced by formaldehyde

Polacow, Ione; Cabasso, Lorrie; Li, Hsueh Jei

doi:10.1021/bi00666a002

Cited by 23 publications

(5 citation statements)

References 41 publications

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“…Formaldehyde or glutaraldehyde has been used (1,6,11,20) to prevent migration and extraction of chromosomal proteins. It should be noted, however, that although exclusive use of cross-linking chemicals may provide additional insight into the organization of proteins in chromosomes, it also creates its own set of artifacts by altering the chromosomal ultrastructure (16,21). Therefore it is expected that the use of several fixation techniques separately but in parallel will provide the most complete information on the organization of chromosomal proteins.…”

Section: Discussionmentioning

confidence: 99%

Histone localization in polytene chromosomes by immunofluorescence.

Kurth¹,

Moudrianakis²,

Bustin³

1978

The Journal of Cell Biology

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Polytene chromosomes of Chironomus thurnmi were treated with antisera elicited by purified calf thymus histone fractions, and the location of each histone type was visualized by the indirect immunofluorescence technique. Each of the antisera produced specific and distinct patterns of fluorescence, suggesting that it is possible to use the indirect immunofluorescence technique to study the in situ organization of each histone in the various regions of the chromosomes. H1 and H2A antisera produced diffuse fluorescence patterns in acetic acid-fixed chromosomes which become more defined in formaldehyde-fixed preparations. Antisera to H2B, H3 and H4, when reacted with either formaldehyde-or acetic acid-fixed chromosomes, produce distinct banding patterns closely resembling the banding of acetoorcein-stained or phase-contrast-differentiated chromosomal preparations. These antisera produce corresponding patterns of fluorescence for each chromosome, suggesting that the overall organization of the histones is similar in the various bands. Because the dense band regions stain more brightly with antihistone sera than the less compacted interband areas, we believe that the number of antigenic sites of chromosome-bound histones is related to the amount of DNA present, and that the accessibility of histone determinants does not differ between the bands and interbands.KEY WORDS histone localization antihistone sera polytene chromosomes 9 Chironomus thummi immunofluorescence Polytene chromosomes provide an excellent system for studying the structural organization of chromosomal constituents because their diameter has been amplified by several consecutive duplications of the basic chromatid fiber without ensuing separation of the sister chromatids. These amplified organelles permit examination not only at the chromosomal level, but also at the level of the gene.Chironomus thummi is an ideal source of these chromosomes because it is easily grown (14) and its chromosomal banding patterns have been well characterized (12). An additional advantage in using Chironornus chromosomes is that they are not fused at the chromocenter as are the chromosomes of Drosophila. This permits easy visualization and analysis of individual chromosomes.The availability of well-characterized antibodies to isolated histones (10, 22) allows the study of the in situ organization of histories within polytene chromosomes. Antihistone sera bind to chromatin (3) and can be used to probe the arrangement of 910 J. CELL BIOLOGY. ~ The Rockefeller University Press 9

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Section: Discussionmentioning

confidence: 99%

Histone localization in polytene chromosomes by immunofluorescence.

Kurth¹,

Moudrianakis²,

Bustin³

1978

The Journal of Cell Biology

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“…The method of using NaCl-treatment of chromatin to obtain partially dehistonized chromatin is suitable. Although histone rearrangement in NaCl solution after formaldehyde fixation has been reported (30), such rearrangement of histones was shown to be caused primarily by reaction of formaldehyde with histones before fixation rather than by NaCl itself (29).…”

Section: Methodsmentioning

confidence: 99%

Structural transition in chromatin induced by ions in solution

Li¹,

Hu²,

Maciewicz³

et al. 1977

Nucl Acids Res

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Structural transition in chromatin was measured as a function of counter ions in solution (NaCl or MgCl(2)) and of histones bound on the DNA. The addition of counter ions to aqueous solutions of chromatin, partially dehistonized chromatin, and DNA caused a drastic reduction in viscosity and a significant increase in sedimentation coefficient. Transitions occurred primarily at about 2 x 10(-3) M NaCl and 1 x 10(-5) M MgCl(2) and are interpreted as a change in structure of chromatin induced by tight binding of cations (Na(+) or Mg(++)) to DNA, either free or bound by histones, and is an intrinsic property of DNA rather than of the type of histone bound. At a given ionic condition, removal of histone H1 from chromatin had only a minor effect on the hydrodynamic properties of chromatin while removal of other histones caused a drastic change in these properties. An increase in the sedimentation coefficient of DNA was observed also for protamine. DNA complexes wherein the bound protein contains only unordered coil rather than the alpha-helices found in histones.

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“…Since the amount of nonreplicated chromatin al ways exceeded the amount of replicated chromatin, most of the newly synthesized protein would associate with nonreplicated chromatin. Fur thermore, exposure of chromatin to fo rmaldehyde can cause redistribution of chromosomal proteins, presumably due to alteration of protein charge (326,327).…”

Section: Distribution Of Old and Ne W His Tones Between Old And Ne W mentioning

confidence: 99%