Mechanisms directing the nuclear localization of the CtBP family proteins

Alexis Verger, Kate G R Quinlan, Linda A Crofts, Stefania Spanò, Daniela Corda, Eleanor P W Kable, Filip Braet, Merlin Crossley

Research output: Contribution to journalArticle

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Abstract

The C-terminal binding protein (CtBP) family includes four proteins (CtBP1 [CtBP1-L], CtBP3/BARS [CtBP1-S], CtBP2, and RIBEYE) which are implicated both in transcriptional repression and in intracellular trafficking. However, the precise mechanisms by which different CtBP proteins are targeted to different subcellular regions remains unknown. Here, we report that the nuclear import of the various CtBP proteins and splice isoforms is differentially regulated. We show that CtBP2 contains a unique nuclear localization signal (NLS) located within its N-terminal region, which contributes to its nuclear accumulation. Using heterokaryon assays, we show that CtBP2 is capable of shuttling between the nucleus and cytoplasm of the cell. Moreover, CtBP2 can heterodimerize with CtBP1-L and CtBP1-S and direct them to the nucleus. This effect strongly depends on the CtBP2 NLS. PXDLS motif-containing transcription factors, such as BKLF, that bind CtBP proteins can also direct them to the nucleus. We also report the identification of a splice isoform of CtBP2, CtBP2-S, that lacks the N-terminal NLS and localizes to the cytoplasm. Finally, we show that mutation of the CtBP NADH binding site impairs the ability of the proteins to dimerize and to associate with BKLF. This reduces the nuclear accumulation of CtBP1. Our results suggest a model in which the nuclear localization of CtBP proteins is influenced by the CtBP2 NLS, by binding to PXDLS motif partner proteins, and through the effect of NADH on CtBP dimerization.
Original languageEnglish
Pages (from-to)4882-94
Number of pages13
JournalMolecular and Cellular Biology
Volume26
Issue number13
DOIs
Publication statusPublished - Jul 2006

Fingerprint

Nuclear Localization Signals
Proteins
NAD
Protein Isoforms
Cytoplasm
Protein Multimerization
Amino Acid Motifs
Cell Nucleus Active Transport
C-terminal binding protein
Cell Nucleus
Protein Binding
Transcription Factors
Binding Sites
Mutation

Keywords

  • Alcohol Oxidoreductases
  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins
  • Carrier Proteins
  • Cell Nucleus
  • Cells, Cultured
  • DNA-Binding Proteins
  • Humans
  • Luminescent Proteins
  • Mice
  • Molecular Sequence Data
  • Nuclear Localization Signals
  • Phosphoproteins
  • Sequence Deletion
  • Transcription Factors

Cite this

Verger, A., Quinlan, K. G. R., Crofts, L. A., Spanò, S., Corda, D., Kable, E. P. W., ... Crossley, M. (2006). Mechanisms directing the nuclear localization of the CtBP family proteins. Molecular and Cellular Biology, 26(13), 4882-94. https://doi.org/10.1128/MCB.02402-05

Mechanisms directing the nuclear localization of the CtBP family proteins. / Verger, Alexis; Quinlan, Kate G R; Crofts, Linda A; Spanò, Stefania; Corda, Daniela; Kable, Eleanor P W; Braet, Filip; Crossley, Merlin.

In: Molecular and Cellular Biology, Vol. 26, No. 13, 07.2006, p. 4882-94.

Research output: Contribution to journalArticle

Verger, A, Quinlan, KGR, Crofts, LA, Spanò, S, Corda, D, Kable, EPW, Braet, F & Crossley, M 2006, 'Mechanisms directing the nuclear localization of the CtBP family proteins', Molecular and Cellular Biology, vol. 26, no. 13, pp. 4882-94. https://doi.org/10.1128/MCB.02402-05
Verger A, Quinlan KGR, Crofts LA, Spanò S, Corda D, Kable EPW et al. Mechanisms directing the nuclear localization of the CtBP family proteins. Molecular and Cellular Biology. 2006 Jul;26(13):4882-94. https://doi.org/10.1128/MCB.02402-05
Verger, Alexis ; Quinlan, Kate G R ; Crofts, Linda A ; Spanò, Stefania ; Corda, Daniela ; Kable, Eleanor P W ; Braet, Filip ; Crossley, Merlin. / Mechanisms directing the nuclear localization of the CtBP family proteins. In: Molecular and Cellular Biology. 2006 ; Vol. 26, No. 13. pp. 4882-94.
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AU - Verger, Alexis

AU - Quinlan, Kate G R

AU - Crofts, Linda A

AU - Spanò, Stefania

AU - Corda, Daniela

AU - Kable, Eleanor P W

AU - Braet, Filip

AU - Crossley, Merlin

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N2 - The C-terminal binding protein (CtBP) family includes four proteins (CtBP1 [CtBP1-L], CtBP3/BARS [CtBP1-S], CtBP2, and RIBEYE) which are implicated both in transcriptional repression and in intracellular trafficking. However, the precise mechanisms by which different CtBP proteins are targeted to different subcellular regions remains unknown. Here, we report that the nuclear import of the various CtBP proteins and splice isoforms is differentially regulated. We show that CtBP2 contains a unique nuclear localization signal (NLS) located within its N-terminal region, which contributes to its nuclear accumulation. Using heterokaryon assays, we show that CtBP2 is capable of shuttling between the nucleus and cytoplasm of the cell. Moreover, CtBP2 can heterodimerize with CtBP1-L and CtBP1-S and direct them to the nucleus. This effect strongly depends on the CtBP2 NLS. PXDLS motif-containing transcription factors, such as BKLF, that bind CtBP proteins can also direct them to the nucleus. We also report the identification of a splice isoform of CtBP2, CtBP2-S, that lacks the N-terminal NLS and localizes to the cytoplasm. Finally, we show that mutation of the CtBP NADH binding site impairs the ability of the proteins to dimerize and to associate with BKLF. This reduces the nuclear accumulation of CtBP1. Our results suggest a model in which the nuclear localization of CtBP proteins is influenced by the CtBP2 NLS, by binding to PXDLS motif partner proteins, and through the effect of NADH on CtBP dimerization.

AB - The C-terminal binding protein (CtBP) family includes four proteins (CtBP1 [CtBP1-L], CtBP3/BARS [CtBP1-S], CtBP2, and RIBEYE) which are implicated both in transcriptional repression and in intracellular trafficking. However, the precise mechanisms by which different CtBP proteins are targeted to different subcellular regions remains unknown. Here, we report that the nuclear import of the various CtBP proteins and splice isoforms is differentially regulated. We show that CtBP2 contains a unique nuclear localization signal (NLS) located within its N-terminal region, which contributes to its nuclear accumulation. Using heterokaryon assays, we show that CtBP2 is capable of shuttling between the nucleus and cytoplasm of the cell. Moreover, CtBP2 can heterodimerize with CtBP1-L and CtBP1-S and direct them to the nucleus. This effect strongly depends on the CtBP2 NLS. PXDLS motif-containing transcription factors, such as BKLF, that bind CtBP proteins can also direct them to the nucleus. We also report the identification of a splice isoform of CtBP2, CtBP2-S, that lacks the N-terminal NLS and localizes to the cytoplasm. Finally, we show that mutation of the CtBP NADH binding site impairs the ability of the proteins to dimerize and to associate with BKLF. This reduces the nuclear accumulation of CtBP1. Our results suggest a model in which the nuclear localization of CtBP proteins is influenced by the CtBP2 NLS, by binding to PXDLS motif partner proteins, and through the effect of NADH on CtBP dimerization.

KW - Alcohol Oxidoreductases

KW - Amino Acid Sequence

KW - Animals

KW - Bacterial Proteins

KW - Carrier Proteins

KW - Cell Nucleus

KW - Cells, Cultured

KW - DNA-Binding Proteins

KW - Humans

KW - Luminescent Proteins

KW - Mice

KW - Molecular Sequence Data

KW - Nuclear Localization Signals

KW - Phosphoproteins

KW - Sequence Deletion

KW - Transcription Factors

U2 - 10.1128/MCB.02402-05

DO - 10.1128/MCB.02402-05

M3 - Article

VL - 26

SP - 4882

EP - 4894

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 13

ER -