DOI: 10.1073/pnas.2035256100. Facile synthesis of site-specifically acetylated and methylated histone proteins: Reagents for evaluation of the histone code hypothesis [Penn]

Facile synthesis of site-specifically acetylated and methylated histone proteins: Reagents for evaluation of the histone code hypothesis

10.1073/pnas.2035256100

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The functional capacity of genetically encoded histone proteins can be powerfully expanded by posttranslational modification. A growing body of biochemical and genetic evidence clearly links the unique combinatorial patterning of side chain acetylation, methylation, and phosphorylation mainly within the highly conserved N termini of histones H2A, H2B, H3, and H4 with the regulation of gene expression and chromatin assembly and remodeling, in effect constituting a “histone code” for epigenetic signaling. Deconvoluting this code has proved challenging given the inherent posttranslational heterogeneity of histone proteins isolated from biological sources. Here we describe the application of native chemical ligation to the preparation of full-length histone proteins containing site-specific acetylation and methylation modifications. Peptide thioesters corresponding to histone N termini were prepared by solid phase peptide synthesis using an acid labile Boc/HF assembly strategy, then subsequently ligated to recombinantly produced histone C-terminal globular domains containing an engineered N-terminal cysteine residue. The ligation site is then rendered traceless by hydrogenolytic desulfurization, generating a native histone protein sequence. Synthetic histones generated by this method are fully functional, as evidenced by their self-assembly into a higher order H3/H4 heterotetramer, their deposition into nucleosomes by human ISWI-containing ( I mitation of Swi tch) factor RSF ( R emodeling and S pacing F actor), and by enzymatic modification by human Sirt1 deacetylase and G9a methyltransferase. Site-specifically modified histone proteins generated by this method will prove invaluable as novel reagents for the evaluation of the histone code hypothesis and analysis of epigenetic signaling mechanisms.

Bibliography

He, S., Bauman, D., Davis, J. S., Loyola, A., Nishioka, K., Gronlund, J. L., Reinberg, D., Meng, F., Kelleher, N., & McCafferty, D. G. (2003). Facile synthesis of site-specifically acetylated and methylated histone proteins: Reagents for evaluation of the histone code hypothesis. Proceedings of the National Academy of Sciences, 100(21), 12033â12038.

Authors 10

Shu He (first)
David Bauman (additional)
Jamaine S. Davis (additional)
Alejandra Loyola (additional)
Kenichi Nishioka (additional)
Jennifer L. Gronlund (additional)
Danny Reinberg (additional)
Fanyu Meng (additional)
Neil Kelleher (additional)
Dewey G. McCafferty (additional)

References 28 Referenced 105

10.1038/nrm1075
10.1016/S0968-0004(00)01718-7
10.1073/pnas.51.5.786
10.1016/S0092-8674(00)00118-5
10.1101/gad.927301
10.1101/gad.12.5.599
10.1016/S0955-0674(02)00335-6
10.1101/gad.937401
10.1002/1521-1878(200009)22:9<836::AID-BIES9>3.0.CO;2-X
10.1038/47412
10.1016/S0092-8674(02)01196-0
10.1111/j.1399-3011.1992.tb00291.x
10.1002/1097-0282(2001)60:3<194::AID-BIP10031>3.0.CO;2-8
10.1016/0003-2697(81)90704-1
Luger, K., Rechsteiner, T. J. & Richmond, T. J. (1999) Methods Mol. Biol. 119, 1–16.10804500 / Methods Mol. Biol. (1999)
10.1021/ja962656r
10.1021/ja003265m
10.1016/S1044-0305(97)00126-8
10.1074/jbc.272.37.23427
10.1101/gad.967202
10.1016/S1097-2765(02)00548-8
10.1126/science.7973629
10.1146/annurev.biochem.69.1.923
10.1021/ja00089a001
10.1073/pnas.91.14.6584
10.1074/jbc.M301445200
10.1126/science.282.5395.1900
Loyola A. He S. Oh S. McCafferty D. G. & Reinberg D. (2003) Methods Enzymol. in press.

Dates

Type	When
Created	21 years, 10 months ago (Oct. 14, 2003, 3:05 p.m.)
Deposited	3 years, 4 months ago (April 25, 2022, 9:34 p.m.)
Indexed	3 weeks, 6 days ago (Aug. 2, 2025, 1:12 a.m.)
Issued	21 years, 10 months ago (Oct. 6, 2003)
Published	21 years, 10 months ago (Oct. 6, 2003)
Published Online	21 years, 10 months ago (Oct. 6, 2003)
Published Print	21 years, 10 months ago (Oct. 14, 2003)

Funders 0

None

BibTeX

@article{He_2003, title={Facile synthesis of site-specifically acetylated and methylated histone proteins: Reagents for evaluation of the histone code hypothesis}, volume={100}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.2035256100}, DOI={10.1073/pnas.2035256100}, number={21}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={He, Shu and Bauman, David and Davis, Jamaine S. and Loyola, Alejandra and Nishioka, Kenichi and Gronlund, Jennifer L. and Reinberg, Danny and Meng, Fanyu and Kelleher, Neil and McCafferty, Dewey G.}, year={2003}, month=oct, pages={12033–12038} }

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