Chromatin associated protease from calf thymus

Dyson, Michael R.; Walker, John M.

doi:10.1111/j.1399-3011.1984.tb00948.x

Cited by 20 publications

(2 citation statements)

References 19 publications

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“…In the context of our findings, the question that arises is whether the truncated HMGB1 exists as a natural protein within the cell or whether it is an artificial product generated solely under the conditions of the tryptic cleavage in vitro. Two lines of evidence support the first possibility: the identification of chromatin-associated trypsin-like protease that cleaves the tail of HMGB1 (36) and the earlier data which documented the presence of a tailless HMGB1 as an endogenous degradation product upon isolation of the protein from calf thymus (37). The acetylation pattern of HMGB1, therefore, might be regarded as a hypothetical in vivo mechanism by which the cell controls some key properties of this protein.…”

Section: Discussionmentioning

confidence: 78%

DNA Bending versus DNA End Joining Activity of HMGB1 Protein Is Modulated in Vitro by Acetylation

et al. 2007

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The ability of HMGB1 protein to recognize bent DNA and to induce bending in linear duplex DNA defines HMGB1 as an architectural factor. It has already been demonstrated that the binding affinity of the protein for various bent DNA structures is enhanced upon in vivo acetylation at Lys2. Here we investigate how this modification of HMGB1 affects its ability to bend DNA. We report that the modified protein cannot bend short DNA fragments but, instead, stimulates joining of the same fragments via their ends. The same properties are exhibited in vivo by acetylated HMGB1 lacking its acidic tail. Further, in vitro acetylation of the truncated protein at Lys81 (possible upon tail removal only) restores the protein's bending ability, while the level of stimulation of DNA end joining is strongly reduced. We conclude, therefore, that the ability of HMGB1 to bend DNA or to stimulate end joining is modulated in vitro by acetylation. In an attempt to explain the properties of in vivo-acetylated HMGB1, its complexes with DNA have been analyzed by both protein-DNA cross-linking and atomic force microscopy. Unlike the parental protein, bound mainly within the internal sequences, acetylated HMGB1 binds preferentially to DNA ends. We propose that the loading of acetylated protein on DNA ends accounts for both the failure to bend DNA and the stimulation of DNA end joining.

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Section: Discussionmentioning

confidence: 78%

DNA Bending versus DNA End Joining Activity of HMGB1 Protein Is Modulated in Vitro by Acetylation

et al. 2007

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“…It should be stressed here, that the truncated tail-less protein should not be regarded solely as an artificial, in vitro generated protein. A trypsin-like chromatin-bound protease has been identified that specifically cleaves the acidic tail of HMGB1 16. Moreover, proteins with truncated C-terminal domain called HMG3 were purified from calf thymus 17 and various tumor cell lines 18.…”

Section: Introductionmentioning

confidence: 99%

The DNA Binding and Bending Activities of Truncated Tail-less HMGB1 protein are Differentially Affected by Lys-2 and Lys-81 Residues and Their Acetylation

Elenkov¹,

Pelovsky²,

Ugrinova³

et al. 2011

Int. J. Biol. Sci.

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The role of lysines 2 and 81 as target sites for acetylation in full-length HMGB1 and truncated tail-less protein, respectively, has been studied by mutation analysis for the abilities of these proteins to bind and bend DNA. The DNA bending ability of truncated tail-less HMGB1 containing Lys-2 mutated to alanine does not differ from that of the wild-type protein, while the same mutation of Lys-81 reduced the bending capacity of the mutant protein. These data demonstrate that Lys-81 is critical for the DNA bending ability of truncated HMGB1. Such a conclusion is further confirmed by the experiments carried out with CBP-acetylated proteins: acetylation of Lys-2 in mutant protein K81/A81 alleviated DNA bending and induced DNA end-joining. On the contrary, the acetylation of Lys-81 in the mutant K2/A2 enhanced the bending potential of HMGB1∆C. Regarding the ability of HMGB1 to specifically bind bent DNA, the individual mutations of either K2 or K81 as well as the double mutation of both residues to alanine were found to completely abolish binding of truncated tail-less HMGB1 to cisplatin-modified DNA. We conclude that unlike the case with the bending ability of truncated HMGB1, where Lys-81 has a primary function, Lys-2 and Lys-81 are both critical for the protein's binding to cisplatin-modified DNA. The mutation K2/A2 in full-length HMGB1 and acidic tail removal induce the same conformational changes. Any further substitutions at the acetylable lysines in the truncated form of HMGB1 do not have an additional effect.

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Identification of a cysteine protease responsible for degradation of sperm histones during male pronucleus remodeling in sea urchins

Imschenetzky

Díaz

Montecino

et al. 1997

J of Cellular Biochemistry

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We have identified a 60-kDa cysteine protease that is associated with chromatin in sea urchin zygotes. This enzyme was found to be present as a proenzyme in unfertilized eggs and was activated shortly after fertilization. At a pH of 7.8-8.0, found after fertilization, the enzyme degraded the five sperm-specific histones (SpH), while the native cleavage-stage (CS) histone variants remained unaffected. Based on its requirements for reducing agents, its inhibition by sulfhydryl blocking compounds and its sensitivity to the cysteine-type protease inhibitors (2S,3S)-trans-epoxysuccinyl-L-leucyl-amido-3-methylbutane-ethyl-es ter (E-64 d), cystatin and leupeptin, this protease can be defined as a cysteine protease. Consistently, this protease was not affected by serine-type protease inhibitors phenylmethylsulfonyl fluoride (PMSF) and pepstatin. The substrate selectivity and pH modulation of the protease activity strongly suggest its role in the removal of sperm-specific histones, which determines sperm chromatin remodeling after fertilization. This suggestion was further substantiated by the inhibition of sperm histones degradation in vivo by E-64 d. Based on these three lines of evidence, we postulate that this cysteine protease is responsible for the degradation of sperm-specific histones which occurs during male pronucleus formation.

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Chromatin associated protease from calf thymus

Cited by 20 publications

References 19 publications

DNA Bending versus DNA End Joining Activity of HMGB1 Protein Is Modulated in Vitro by Acetylation

DNA Bending versus DNA End Joining Activity of HMGB1 Protein Is Modulated in Vitro by Acetylation

The DNA Binding and Bending Activities of Truncated Tail-less HMGB1 protein are Differentially Affected by Lys-2 and Lys-81 Residues and Their Acetylation

Identification of a cysteine protease responsible for degradation of sperm histones during male pronucleus remodeling in sea urchins

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