Further Studies of a Thymus Nucleohistone-associated Protease

Bartley, James; Chalkley, Roger

doi:10.1016/s0021-9258(19)63792-0

Cited by 180 publications

(20 citation statements)

References 19 publications

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“…Histories were prepared from calfthymus tissue, which had been frozen in liquid nitrogen immediately after slaughtering. Nucleohistone was purified as described by Johns (4) using 50 mM sodium hydrogensulfit, pH 7.4, in 0.1 M sodium chloride as a proteolysis inhibitor (45). Total histone was extracted from nucleohistone with 0.2 M sulfuric acid, precipitated in ethanol, and lyophilized.…”

Section: Methodsmentioning

confidence: 99%

Histones have high affinity for the glomerular basement membrane. Relevance for immune complex formation in lupus nephritis.

Schmiedeke

Stockl

Weber

et al. 1989

The Journal of Experimental Medicine

141

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Histones are a set of highly cationic proteins essentially involved in the binding and compaction of DNA in the cell nucleus chromatin (1-3). In mammals, five subclasses of histones exist: H1 (f1), H2a (f2a2), H2b (f2b), H3 (f3), and H4 (f2a1) (4,5), whereby four histones form an octameric core protein aggregate (H2a, H2b, H3, H4) around which a 145-bp DNA molecule is wrapped, forming the core particle followed by a variable (-55 bp) DNA region, complexed with H1 (6). In the cell nucleus, histones and DNA are present in similar quantities .These components of the cell nucleus are two of the most important antigens for autoantibody formation in SLE (7-12), and it has been reported that the titers of both groups of autoantibodies correlate well with disease activity (13-16).A prominent manifestation oforgan involvement in SLE patients is the occurrence of immune complex (IC)' glomerulonephritis . After the original report by Koffler et al. (17), attention has been focussed on the role of DNA-anti-DNA IC for >20 yr. Two proposals have been widely discussed in connection with the glomerular deposition of DNA-anti-DNA complexes: the now unfashionable notion that preformed soluble ICs can locate in the glomerular filter (18); and second, an affinity of DNA for the glomerular basement membrane (GBM) in vivo has been suggested (19-23), which could then initiate in situ IC formation. Extensive studies in animals on the kinetics and clearance of circulating free or immune complexed DNA (24-27) in regard to size and strandedness of DNA, however, revealed rapid nonrenal uptake and clearance. Prolonged DNA persistence is difficult to explain by accepted concepts; new proposals are needed (28). Contradictory findings on the occurrence of free or IC-bound DNA in sera from normal individuals and SLE patients have been reported (for review see reference 29). There are a number of technical problems involved here and it is now accepted that DNA-anti-DNA immune complexes represent only a minor part of the ICs found in lupus patients, and that increased levels ofcirculating DNA are found in a variety of conditions in which lysis ofcells occurs . Preliminary studies by Fournie (29) suggest that nucleosomes, which consist ofDNA This work was supported by the Deutsche Forschungsgemeinschaft, Ba 907/1-1.

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Section: Methodsmentioning

confidence: 99%

Histones have high affinity for the glomerular basement membrane. Relevance for immune complex formation in lupus nephritis.

Schmiedeke

Stockl

Weber

et al. 1989

The Journal of Experimental Medicine

141

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“…The nature of histone fraction Fl and its digestion by endogenous protease will be the subject of another communication . Similar F1-specific proteolysis has been described in calf thymus chromatin (Panyim et al ., 1968 ;Bartley and Chalkley, 1970) . Histones were extracted from calf thymus chromatin by the method of .…”

Section: Extraction Of Histonementioning

confidence: 99%

“…1 a shows a densitometer tracing of histones isolated from macronuclei . Since Tetrahymena contains a histone protease with activity similar to that described in calf thymus (Bartley and Chalkley, 1970), we have routinely removed fraction F1 and its digestion products with perchloric acid so that they do not interfere with the visualization of fraction F2A1 . The other electrophoretically separable fractions of Tetrahymena histones have been identified by the electrophoretic criteria of and by differential solubilization using the fractionation criteria described by Johns (1964Johns ( , 1967 .…”

Section: Urea-acrylamide Gels Of Tetrahymena Histonesmentioning

confidence: 99%

STUDIES ON HISTONE FRACTION F2A1 IN MACRO- AND MICRONUCLEI OF TETRAHYMENA PYRIFORMIS

Gorovsky

Pleger

Keevert

et al. 1973

The Journal of Cell Biology

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Histone fraction F2A1 has been isolated and purified from macronuclei of the ciliate Tetrahymena pyriformis . It migrates as a single species on sodium dodecyl sulphate-acrylamide gel electrophoresis, with a molecular weight indistinguishable from that of calf thymus F2A1 . The solubility properties of Tetrahymena F2AI are also similar to those of calf thymus F2A1 . Electrophoretic analyses on urea-acrylamide gels indicate that Tetrahymena F2A1 consists of four or five subspecies, the two fastest having electrophoretic mobilities identical with those of the two major electrophoretically separable forms of calf thymus F2A1 . High resolution (long gel) electrophoresis coupled with incorporation of radioactive acetate both in vivo and in vitro suggest that, as in the case of calf thymus F2A1, differentical acetylation of a parent molecule can explain the observed electrophoretic heterogeneity of Tetrahymena F2A1 . Electrophoretic analysis of histones isolated from the micronucleus, which is genetically less active than the macronucleus, indicates that it contains largely the relatively unacetylated (parent) form of histone F2A1 .

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“…The reason for this discrepancy is unclear . nuclei and histones were isolated in 0.05 M bisulfite to inhibit proteolysis (Panyim et al, 1968 ;Bartley and Chalkley, 1970) . Moreover, the relative amounts of all three Fl bands were found to be constant (Table II), suggesting that the minor bands are not the result of degradation .…”

Section: Identification and Characterization Of Histone Fractionsmentioning

confidence: 99%

The Histones Associated With Condensed and Extended Chromatin of Mouse Liver

Johmann

Eckhardt

Gorovsky

1973

The Journal of Cell Biology

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Histones were extracted from isolated mouse liver nuclei, and from mouse liver condensed and extended chromatin . Mouse liver histones were found to be very similar to those of calf thymus in their solubility properties, relative electrophoretic mobilities, and molecular weights as determined on SDS-polyacrylamide gels . Quantitative analysis by high-resolution gel electrophoresis demonstrated a remarkable similarity between the histones of condensed chromatin and those of extended chromatin . However, minor differences were found . A unique subspecies was found only in condensed chromatin histone and the relative amounts of fractions F2AI and F2A2 differed in the two types of chromatin . The ratio of the parental to the acetylated form of F2A1 was identical in the two chromatin samples . Since DNA extracted from the condensed chromatin fraction consisted of approximately 50% satellite DNA, the general similarities between the histones of condensed and extended chromatin make it likely that even this simple, highly repetitive DNA is complexed with a number of histone subfractions .

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Further Studies of a Thymus Nucleohistone-associated Protease

Cited by 180 publications

References 19 publications

Histones have high affinity for the glomerular basement membrane. Relevance for immune complex formation in lupus nephritis.

Histones have high affinity for the glomerular basement membrane. Relevance for immune complex formation in lupus nephritis.

STUDIES ON HISTONE FRACTION F2A1 IN MACRO- AND MICRONUCLEI OF TETRAHYMENA PYRIFORMIS

The Histones Associated With Condensed and Extended Chromatin of Mouse Liver

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