title: Phosphorylation of Krüppel-like factor 3 (KLF3/BKLF) and C-terminal binding protein 2 (CtBP2) by homeodomain-interacting protein kinase 2 (HIPK2) modulates KLF3 DNA binding and activity.
abstract: Krüppel-like factor 3 (KLF3/BKLF), a member of the Krüppel-like factor (KLF) family of transcription factors, is a widely expressed transcriptional repressor with diverse biological roles. Although there is considerable understanding of the molecular mechanisms that allow KLF3 to silence the activity of its target genes, less is known about the signal transduction pathways and post-translational modifications that modulate KLF3 activity in response to physiological stimuli. We observed that KLF3 is modified in a range of different tissues and found that the serine/threonine kinase homeodomain-interacting protein kinase 2 (HIPK2) can both bind and phosphorylate KLF3. Mass spectrometry identified serine 249 as the primary phosphorylation site. Mutation of this site reduces the ability of KLF3 to bind DNA and repress transcription. Furthermore, we also determined that HIPK2 can phosphorylate the KLF3 co-repressor C-terminal binding protein 2 (CtBP2) at serine 428. Finally, we found that phosphorylation of KLF3 and CtBP2 by HIPK2 strengthens the interaction between these two factors and increases transcriptional repression by KLF3. Taken together, our results indicate that HIPK2 potentiates the activity of KLF3.
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Vitri
authors: V
authors: Dewi
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Alister
authors: A
authors: Kwok
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Stella
authors: S
authors: Lee
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Ming Min
authors: MM
authors: Lee
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Yee Mun
authors: YM
authors: Tan
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Hannah R
authors: HR
authors: Nicholas
authors: the RIKEN Research Center for Allergy and Immunology, Tsurumi-ku, Yokohama City, Kanagawa 230-0045, Japan.
authors: Kyo-ichi
authors: K
authors: Isono
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Beeke
authors: B
authors: Wienert
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Ka Sin
authors: KS
authors: Mak
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Alexander J
authors: AJ
authors: Knights
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Kate G R
authors: KG
authors: Quinlan
authors: the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and.
authors: Stuart J
authors: SJ
authors: Cordwell
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Alister P W
authors: AP
authors: Funnell
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors: Richard C M
authors: RC
authors: Pearson
authors: From the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia, the School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia, and m.crossley@unsw.edu.au.
authors: Merlin
authors: M
authors: Crossley
mesh_terms: D000429:Alcohol Oxidoreductases; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000502:physiology*; D002522:Chlorocebus aethiops; D056970:Co-Repressor Proteins; D004247:DNA / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D051741:Kruppel-Like Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D002352:Carrier Proteins; D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins; C529780:Klf3 protein, mouse; D051741:Kruppel-Like Transcription Factors; D010750:Phosphoproteins; D004247:DNA; D000429:Alcohol Oxidoreductases; C111341:Ctbp2 protein, mouse; C492003:Hipk2 protein, mouse; D017346:Protein Serine-Threonine Kinases
keywords: C-terminal Binding Protein (CtBP2); Gene Expression; Homeodomain-interacting Protein Kinase 2 (HIPK2); Kruppel-like Factor (KLF); Kruppel-like Factor 3 (KLF3/BKLF); Post-translational Modification (PTM); Protein Phosphorylation; Transcription Regulation