Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1

George Hoppe, Katherine E. Talcott, Sanjoy K Bhattacharya, John W. Crabb, Jonathan E. Sears

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Oxidative stress can induce a covalent disulfide bond between protein and peptide thiols that is reversible through enzymatic catalysis. This process provides a post-translational mechanism for control of protein function and may also protect thiol groups from irreversible oxidation. High mobility group protein B1 (Hmgb1), a DNA-binding structural chromosomal protein and transcriptional co-activator was identified as a substrate of glutaredoxin. Hmgb1 contains 3 cysteines, Cys23, 45, and 106. In mild oxidative conditions, Cys23 and Cys45 readily form an intramolecular disulfide bridge, whereas Cys106 remains in the reduced form. The disulfide bond between Cys23 and Cys45 is a target of glutathione-dependent reduction by glutaredoxin. Endogenous Hmgb1 as well as GFP-tagged wild-type Hmgb1 co-localize in the nucleus of CHO cells. While replacement of Hmgb1 Cys23 and/or 45 with serines did not affect the nuclear distribution of the mutant proteins, Cys106-to-Ser and triple cysteine mutations impaired nuclear localization of Hmgb1. Our cysteine targeted mutational analysis suggests that Cys23 and 45 induce conformational changes in response to oxidative stress, whereas Cys106 appears to be critical for the nucleocytoplasmic shuttling of Hmgb1.

Original languageEnglish
Pages (from-to)3526-3538
Number of pages13
JournalExperimental Cell Research
Volume312
Issue number18
DOIs
StatePublished - Nov 1 2006
Externally publishedYes

Fingerprint

High Mobility Group Proteins
Cell Nucleus Active Transport
Chromatin
Oxidation-Reduction
Glutaredoxins
Disulfides
Proteins
Cysteine
Sulfhydryl Compounds
Oxidative Stress
CHO Cells
Mutant Proteins
Catalysis
Serine
Glutathione
Peptides
Mutation
DNA

Keywords

  • Chromatin remodeling
  • Cysteine
  • GFP
  • Glutaredoxin
  • High mobility group
  • Oxidative stress
  • Thiol-disulfide reactions

ASJC Scopus subject areas

  • Cell Biology

Cite this

Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1. / Hoppe, George; Talcott, Katherine E.; Bhattacharya, Sanjoy K; Crabb, John W.; Sears, Jonathan E.

In: Experimental Cell Research, Vol. 312, No. 18, 01.11.2006, p. 3526-3538.

Research output: Contribution to journalArticle

Hoppe, George ; Talcott, Katherine E. ; Bhattacharya, Sanjoy K ; Crabb, John W. ; Sears, Jonathan E. / Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1. In: Experimental Cell Research. 2006 ; Vol. 312, No. 18. pp. 3526-3538.
@article{f6b5f297aa084fd6a54ccbbe41697052,
title = "Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1",
abstract = "Oxidative stress can induce a covalent disulfide bond between protein and peptide thiols that is reversible through enzymatic catalysis. This process provides a post-translational mechanism for control of protein function and may also protect thiol groups from irreversible oxidation. High mobility group protein B1 (Hmgb1), a DNA-binding structural chromosomal protein and transcriptional co-activator was identified as a substrate of glutaredoxin. Hmgb1 contains 3 cysteines, Cys23, 45, and 106. In mild oxidative conditions, Cys23 and Cys45 readily form an intramolecular disulfide bridge, whereas Cys106 remains in the reduced form. The disulfide bond between Cys23 and Cys45 is a target of glutathione-dependent reduction by glutaredoxin. Endogenous Hmgb1 as well as GFP-tagged wild-type Hmgb1 co-localize in the nucleus of CHO cells. While replacement of Hmgb1 Cys23 and/or 45 with serines did not affect the nuclear distribution of the mutant proteins, Cys106-to-Ser and triple cysteine mutations impaired nuclear localization of Hmgb1. Our cysteine targeted mutational analysis suggests that Cys23 and 45 induce conformational changes in response to oxidative stress, whereas Cys106 appears to be critical for the nucleocytoplasmic shuttling of Hmgb1.",
keywords = "Chromatin remodeling, Cysteine, GFP, Glutaredoxin, High mobility group, Oxidative stress, Thiol-disulfide reactions",
author = "George Hoppe and Talcott, {Katherine E.} and Bhattacharya, {Sanjoy K} and Crabb, {John W.} and Sears, {Jonathan E.}",
year = "2006",
month = "11",
day = "1",
doi = "10.1016/j.yexcr.2006.07.020",
language = "English",
volume = "312",
pages = "3526--3538",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "18",

}

TY - JOUR

T1 - Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1

AU - Hoppe, George

AU - Talcott, Katherine E.

AU - Bhattacharya, Sanjoy K

AU - Crabb, John W.

AU - Sears, Jonathan E.

PY - 2006/11/1

Y1 - 2006/11/1

N2 - Oxidative stress can induce a covalent disulfide bond between protein and peptide thiols that is reversible through enzymatic catalysis. This process provides a post-translational mechanism for control of protein function and may also protect thiol groups from irreversible oxidation. High mobility group protein B1 (Hmgb1), a DNA-binding structural chromosomal protein and transcriptional co-activator was identified as a substrate of glutaredoxin. Hmgb1 contains 3 cysteines, Cys23, 45, and 106. In mild oxidative conditions, Cys23 and Cys45 readily form an intramolecular disulfide bridge, whereas Cys106 remains in the reduced form. The disulfide bond between Cys23 and Cys45 is a target of glutathione-dependent reduction by glutaredoxin. Endogenous Hmgb1 as well as GFP-tagged wild-type Hmgb1 co-localize in the nucleus of CHO cells. While replacement of Hmgb1 Cys23 and/or 45 with serines did not affect the nuclear distribution of the mutant proteins, Cys106-to-Ser and triple cysteine mutations impaired nuclear localization of Hmgb1. Our cysteine targeted mutational analysis suggests that Cys23 and 45 induce conformational changes in response to oxidative stress, whereas Cys106 appears to be critical for the nucleocytoplasmic shuttling of Hmgb1.

AB - Oxidative stress can induce a covalent disulfide bond between protein and peptide thiols that is reversible through enzymatic catalysis. This process provides a post-translational mechanism for control of protein function and may also protect thiol groups from irreversible oxidation. High mobility group protein B1 (Hmgb1), a DNA-binding structural chromosomal protein and transcriptional co-activator was identified as a substrate of glutaredoxin. Hmgb1 contains 3 cysteines, Cys23, 45, and 106. In mild oxidative conditions, Cys23 and Cys45 readily form an intramolecular disulfide bridge, whereas Cys106 remains in the reduced form. The disulfide bond between Cys23 and Cys45 is a target of glutathione-dependent reduction by glutaredoxin. Endogenous Hmgb1 as well as GFP-tagged wild-type Hmgb1 co-localize in the nucleus of CHO cells. While replacement of Hmgb1 Cys23 and/or 45 with serines did not affect the nuclear distribution of the mutant proteins, Cys106-to-Ser and triple cysteine mutations impaired nuclear localization of Hmgb1. Our cysteine targeted mutational analysis suggests that Cys23 and 45 induce conformational changes in response to oxidative stress, whereas Cys106 appears to be critical for the nucleocytoplasmic shuttling of Hmgb1.

KW - Chromatin remodeling

KW - Cysteine

KW - GFP

KW - Glutaredoxin

KW - High mobility group

KW - Oxidative stress

KW - Thiol-disulfide reactions

UR - http://www.scopus.com/inward/record.url?scp=33749125188&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749125188&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2006.07.020

DO - 10.1016/j.yexcr.2006.07.020

M3 - Article

C2 - 16962095

AN - SCOPUS:33749125188

VL - 312

SP - 3526

EP - 3538

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 18

ER -