A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress

Victor M. Brown, Eugene Y. Krynetski, Natalia F. Krynetskaia, Dara Grieger, Suraj T. Mukatira, Kuruganti G. Murti, Clive A. Slaughter, Hee Won Park, William E. Evans

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein with glycolytic and non-glycolytic functions, including pro-apoptotic activity. GAPDH accumulates in the nucleus after cells are treated with genotoxic drugs, and it is present in a protein complex that binds DNA modified by thioguanine incorporation. We identified a novel CRM1-dependent nuclear export signal (NES) comprising 13 amino acids (KKVVKQASEGPLK) in the C-terminal domain of GAPDH, truncation or mutation of which abrogated CRM1 binding and caused nuclear accumulation of GAPDH. Alanine scanning of the sequence encompassing the putative NES demonstrated at least two regions important for nuclear export. Site mutagenesis of Lys259 did not affect oligomerization but impaired nuclear efflux of GAPDH, indicating that this amino acid residue is essential for proper functioning of this NES. This novel NES does not contain multiple leucine residues unlike other CRM1-interacting NES, is conserved in GAPDH from multiple species, and has sequence similarities to the export signal found in feline immunodeficiency virus Rev protein. Similar sequences (KKVV*7-13PLK) were found in two other human proteins, U5 small nuclear ribonucleoprotein, and transcription factor BT3.

Original languageEnglish (US)
Pages (from-to)5984-5992
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number7
DOIs
StatePublished - Feb 13 2004
Externally publishedYes

Fingerprint

Nuclear Export Signals
Glyceraldehyde-3-Phosphate Dehydrogenases
DNA Damage
U5 Small Nuclear Ribonucleoproteins
rev Gene Products
Feline Immunodeficiency Virus
Thioguanine
Amino Acids
Oligomerization
Mutagenesis
Proteins
Cell Nucleus Active Transport
Cell Nucleus
Viruses
Leucine
Alanine
Transcription Factors
Scanning
Mutation
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Brown, V. M., Krynetski, E. Y., Krynetskaia, N. F., Grieger, D., Mukatira, S. T., Murti, K. G., ... Evans, W. E. (2004). A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress. Journal of Biological Chemistry, 279(7), 5984-5992. https://doi.org/10.1074/jbc.M307071200

A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress. / Brown, Victor M.; Krynetski, Eugene Y.; Krynetskaia, Natalia F.; Grieger, Dara; Mukatira, Suraj T.; Murti, Kuruganti G.; Slaughter, Clive A.; Park, Hee Won; Evans, William E.

In: Journal of Biological Chemistry, Vol. 279, No. 7, 13.02.2004, p. 5984-5992.

Research output: Contribution to journalArticle

Brown, Victor M. ; Krynetski, Eugene Y. ; Krynetskaia, Natalia F. ; Grieger, Dara ; Mukatira, Suraj T. ; Murti, Kuruganti G. ; Slaughter, Clive A. ; Park, Hee Won ; Evans, William E. / A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 7. pp. 5984-5992.
@article{ad257ba6a94d4061a2e21718a28657cf,
title = "A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress",
abstract = "Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein with glycolytic and non-glycolytic functions, including pro-apoptotic activity. GAPDH accumulates in the nucleus after cells are treated with genotoxic drugs, and it is present in a protein complex that binds DNA modified by thioguanine incorporation. We identified a novel CRM1-dependent nuclear export signal (NES) comprising 13 amino acids (KKVVKQASEGPLK) in the C-terminal domain of GAPDH, truncation or mutation of which abrogated CRM1 binding and caused nuclear accumulation of GAPDH. Alanine scanning of the sequence encompassing the putative NES demonstrated at least two regions important for nuclear export. Site mutagenesis of Lys259 did not affect oligomerization but impaired nuclear efflux of GAPDH, indicating that this amino acid residue is essential for proper functioning of this NES. This novel NES does not contain multiple leucine residues unlike other CRM1-interacting NES, is conserved in GAPDH from multiple species, and has sequence similarities to the export signal found in feline immunodeficiency virus Rev protein. Similar sequences (KKVV*7-13PLK) were found in two other human proteins, U5 small nuclear ribonucleoprotein, and transcription factor BT3.",
author = "Brown, {Victor M.} and Krynetski, {Eugene Y.} and Krynetskaia, {Natalia F.} and Dara Grieger and Mukatira, {Suraj T.} and Murti, {Kuruganti G.} and Slaughter, {Clive A.} and Park, {Hee Won} and Evans, {William E.}",
year = "2004",
month = "2",
day = "13",
doi = "10.1074/jbc.M307071200",
language = "English (US)",
volume = "279",
pages = "5984--5992",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "7",

}

TY - JOUR

T1 - A Novel CRM1-mediated Nuclear Export Signal Governs Nuclear Accumulation of Glyceraldehyde-3-phosphate Dehydrogenase following Genotoxic Stress

AU - Brown, Victor M.

AU - Krynetski, Eugene Y.

AU - Krynetskaia, Natalia F.

AU - Grieger, Dara

AU - Mukatira, Suraj T.

AU - Murti, Kuruganti G.

AU - Slaughter, Clive A.

AU - Park, Hee Won

AU - Evans, William E.

PY - 2004/2/13

Y1 - 2004/2/13

N2 - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein with glycolytic and non-glycolytic functions, including pro-apoptotic activity. GAPDH accumulates in the nucleus after cells are treated with genotoxic drugs, and it is present in a protein complex that binds DNA modified by thioguanine incorporation. We identified a novel CRM1-dependent nuclear export signal (NES) comprising 13 amino acids (KKVVKQASEGPLK) in the C-terminal domain of GAPDH, truncation or mutation of which abrogated CRM1 binding and caused nuclear accumulation of GAPDH. Alanine scanning of the sequence encompassing the putative NES demonstrated at least two regions important for nuclear export. Site mutagenesis of Lys259 did not affect oligomerization but impaired nuclear efflux of GAPDH, indicating that this amino acid residue is essential for proper functioning of this NES. This novel NES does not contain multiple leucine residues unlike other CRM1-interacting NES, is conserved in GAPDH from multiple species, and has sequence similarities to the export signal found in feline immunodeficiency virus Rev protein. Similar sequences (KKVV*7-13PLK) were found in two other human proteins, U5 small nuclear ribonucleoprotein, and transcription factor BT3.

AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein with glycolytic and non-glycolytic functions, including pro-apoptotic activity. GAPDH accumulates in the nucleus after cells are treated with genotoxic drugs, and it is present in a protein complex that binds DNA modified by thioguanine incorporation. We identified a novel CRM1-dependent nuclear export signal (NES) comprising 13 amino acids (KKVVKQASEGPLK) in the C-terminal domain of GAPDH, truncation or mutation of which abrogated CRM1 binding and caused nuclear accumulation of GAPDH. Alanine scanning of the sequence encompassing the putative NES demonstrated at least two regions important for nuclear export. Site mutagenesis of Lys259 did not affect oligomerization but impaired nuclear efflux of GAPDH, indicating that this amino acid residue is essential for proper functioning of this NES. This novel NES does not contain multiple leucine residues unlike other CRM1-interacting NES, is conserved in GAPDH from multiple species, and has sequence similarities to the export signal found in feline immunodeficiency virus Rev protein. Similar sequences (KKVV*7-13PLK) were found in two other human proteins, U5 small nuclear ribonucleoprotein, and transcription factor BT3.

UR - http://www.scopus.com/inward/record.url?scp=1242316968&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1242316968&partnerID=8YFLogxK

U2 - 10.1074/jbc.M307071200

DO - 10.1074/jbc.M307071200

M3 - Article

VL - 279

SP - 5984

EP - 5992

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 7

ER -