A Phosphomimetic Study Implicates Ser557 in Regulation of FOXP2 DNA Binding

A Phosphomimetic Study Implicates Ser557 in Regulation of FOXP2 DNA Binding FOXP2 is a transcription factor expressed in multiple tissues during embryonic development. FOXP2 regulates transcription by binding to DNA at its DNA binding domain, the forkhead domain (FHD) through the recognition helix. Ser557 is a residue located within the recognition helix that has the potential to become phosphorylated posttranslationally. In this study we investigated whether phosphorylation of Ser557 can influence the structure and DNA binding of the FOXP2 FHD. We did this by constructing S557E, a phosphomimetic mutant, and comparing its behaviour to the wild type. The mutation did not affect the secondary or tertiary structure of the protein although it did decrease the propensity of the FOXP2 FHD to form dimers. Most notably, the mutation showed significantly reduced DNA binding compared to the wild type as detected using electrophoretic mobility shift assays. Molecular docking was also performed in which the wild type, phosphomimetic mutant and phosphorylated wild-type were docked to DNA and their interactions with DNA were compared. These results indicated that the wild type forms more interactions with the DNA and that the phosphomimetic mutant as well as the phosphorylated wild type did not associate as favourably with the DNA. This indicates that phosphorylation of Ser557 could disrupt DNA binding likely due to electrostatic and steric hindrance. This suggests that phosphorylation of Ser557 in the FOXP2 FHD could act as a control mechanism for FOXP2 and ultimately could be involved in regulation of transcription. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Protein Journal Springer Journals

A Phosphomimetic Study Implicates Ser557 in Regulation of FOXP2 DNA Binding

13 pages
Read Article

Loading next page...
 
/lp/springer_journal/a-phosphomimetic-study-implicates-ser557-in-regulation-of-foxp2-dna-bdEosOToGa
Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer Science+Business Media, LLC, part of Springer Nature
Subject
Chemistry; Bioorganic Chemistry; Biochemistry, general; Organic Chemistry; Animal Anatomy / Morphology / Histology
ISSN
1572-3887
eISSN
1573-4943
D.O.I.
10.1007/s10930-018-9777-7
Publisher site
See Article on Publisher Site

Abstract

FOXP2 is a transcription factor expressed in multiple tissues during embryonic development. FOXP2 regulates transcription by binding to DNA at its DNA binding domain, the forkhead domain (FHD) through the recognition helix. Ser557 is a residue located within the recognition helix that has the potential to become phosphorylated posttranslationally. In this study we investigated whether phosphorylation of Ser557 can influence the structure and DNA binding of the FOXP2 FHD. We did this by constructing S557E, a phosphomimetic mutant, and comparing its behaviour to the wild type. The mutation did not affect the secondary or tertiary structure of the protein although it did decrease the propensity of the FOXP2 FHD to form dimers. Most notably, the mutation showed significantly reduced DNA binding compared to the wild type as detected using electrophoretic mobility shift assays. Molecular docking was also performed in which the wild type, phosphomimetic mutant and phosphorylated wild-type were docked to DNA and their interactions with DNA were compared. These results indicated that the wild type forms more interactions with the DNA and that the phosphomimetic mutant as well as the phosphorylated wild type did not associate as favourably with the DNA. This indicates that phosphorylation of Ser557 could disrupt DNA binding likely due to electrostatic and steric hindrance. This suggests that phosphorylation of Ser557 in the FOXP2 FHD could act as a control mechanism for FOXP2 and ultimately could be involved in regulation of transcription.

Journal

The Protein Journal – Springer Journals

Published: May 29, 2018

References

  • Prokaryotic transcription regulators: more than just the helix-turn-helix motif
    Huffman, JL; Brennan, RG
  • Transcription factors: an overview
    Latchman, S
  • A post-translational modification code for transcription factors: sorting through a sea of signals
    Benayoun, BA; Veitia, RA
  • The effects of phosphorylation on the structure and function of proteins
    Johnson, L; Barford, D
  • Post-translational modifications influence transcription factor activity: a view from the ETS superfamily
    Tootle, TL; Rebay, I
  • Conserved mechanisms of Ras regulation of evolutionary related transcription factors, Ets1 and Pointed P2
    Wasylyk, C; Bradford, A; Gutierrez-Hartmann, A; Wasylyk, B
  • IL-6-regulated transcription factors
    Akira, S
  • The Foxa family of transcription factors in development and metabolism
    Friedman, JR; Kaestner, KH
  • Co-crystal structure of the HNF-3/fork head DNA-recognition motif resembles histone H5
    Clark, K; Halay, E; Lai, E; Burley, S
  • Multiple domains define the expression and regulatory properties of Foxp1 forkhead transcriptional repressors
    Wang, B; Lin, D; Li, C; Tucker, P
  • The FOXP1 winged helix transcription factor is a novel candidate tumor suppressor gene on chromosome 3p1
    Banham, AH; Beasley, N; Campo, E
  • Domain requirements and sequence specificity of DNA binding for the forkhead transcription factor FOXP3
    Koh, KP; Sundrud, MS; Rao, A
  • FOXP2 as a molecular window into speech and language
    Fisher, SE; Scharff, C
  • Genetic variation in FOXP2 alters grey matter concentrations in schizophrenia patients
    Spaniel, FJ; Horacek, J; Tintera, I; Ibrahim, T; Novak, J; Cermak, M; Klirova; Hoschl, C
  • The evolution of Fox genes and their role in development and disease
    Hannenhalli, S; Kaestner, KH
  • The immune dysregulation polyendocrinopathy enteropathy x-linked syndrome (IPEX) is caused by mutations in FOXP3
    Bennett, CL; Christie, J; Ramsdell, F; Brunkow, ME; Ferguson, PJ; Witesell, L
  • Association between the FOXP2 gene and autistic disorder in Chinese population
    Gong, X; Jia, M; Ruan, Y; Shuang, M; Liu, J; Wu, S; Guo, Y; Yang, J; Ling, Y; Yang, X; Zhang, D
  • Control of Huntington’s disease-associated phenotypes by the striatum enriched transcription factor Foxp2
    Hachigian, LJ; Carmona, V; Fenster, RJ; Kulicke, R; Heilbut, A; Sittler, A; Almeida, P; Mesirov, L; Gao, JP; Kolaczyk, F; Heiman, M
  • Transcriptional and DNA binding activity of the Foxp1/2/4 family is modulated by heterotypic and homotypic protein interactions
    Li, S; Weidenfeld, J; Morrisey, EE
  • Structure of a domain-swapped FOXP3 Dimer on DNA and its function in regulatory T cells
    Bandukwala, HS; Wu, Y; Feuerer, M
  • Structure of the forkhead domain of FOXP2 bound to DNA
    Stroud, JC; Wu, Y; Bates, DL
  • Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1
    Biggs, WH; Meisenhelder, J; Hunter, T
  • Phosphorylation of serine 256 suppresses transactivation by FKHR (FOXO1) by multiple mechanisms: Direct and indirect effects on nuclear/cytoplasmic shuttling and DNA binding
    Zhang, X; Gan, L; Pan, H
  • Structure/function relationships underlying regulation of FOXO transcription factors
    Obsil, T; Obsilova, V
  • Phosphorylation of FOXP3 controls regulatory T cell function and is inhibited by TNF-α in rheumatoid arthritis
    Nie, H; Zheng, Y; Li, R
  • The establishment of a predictive mutational model of the forkhead domain through the analyses of FOXC2 missense mutations identified in patients with hereditary lymphedema with distichiasis
    Berry, FB; Tamimi, Y; Carle, MV
  • A conserved phosphorylation site within the forkhead domain of FoxM1B is required for its activation by cyclin-CDK1
    Chen, YJ; Dominguez-Brauer, C; Wang, Z
  • Identification of missing proteins in the neXtProt database and unregistered phosphopeptides in the phosphositeplus database as part of the chromosome-centric human proteome project
    Shiromizu, T; Adachi, J; Watanabe, S
  • A Conserved MST-FOXO signaling pathway mediates oxidative-stress responses and extends life span
    Lehtinen, MK; Yuan, Z; Boag, PR
  • Structural basis for DNA recognition by FoxO1 and its regulation by posttranslational modification
    Brent, MM; Anand, R; Marmorstein, R
  • Mutation of phosphoserine 389 affects p53 function in vivo
    Lowy, AM
  • Phosphorylation of the kinase homology domain is essential for activation of the A-type natriuretic peptide receptor
    Potter, LR; Hunter, T
  • The chemical biology of protein phosphorylation
    Tarrant, MK; Cole, PA
  • Effect of pH on the structure and DNA binding of the FOXP2 forkhead domain
    Blane, A; Fanucchi, S
  • Microfluidic affinity and ChIP-seq analyses converge on a conserved FOXP2-binding motif in chimp and human, which enables the detection of evolutionarily novel targets
    Nelson, CS; Fuller, CK; Fordyce, PM
  • The FOXP2 forkhead domain binds to a variety of DNA sequences with different rates and affinities
    Webb, H; Steeb, O; Blane, A
  • OligoCalc: an online oligonucleotide properties calculator
    Kibbe, WA
  • A single amino acid in the hinge loop region of the FOXP Forkhead domain is significant for dimerisation
    Perumal, K; Dirr, HW; Fanucchi, S
  • UCSF Chimera: a visualization system for exploratory research and analysis
    Pettersen, EF; Goddard, TD; Huang, CC
  • HADDOCK: a protein-protein docking approach based on biochemical or biophysical information
    Dominguez, C; Boelens, R; Bonvin, AMJJ
  • MolProbity: all-atom structure validation for macromolecular crystallography
    Chen, VB; Arendall, WB; Headd, JJ; Keedy, DA; Immormino, RM; Kapral, GJ; Murray, LW; Richardson, JS; Richardson, DC
  • MolProbity: all-atom contacts and structure validation for proteins and nucleic acids
    Davis, IW; Leaver-Fay, A; Chen, VB; Block, JN; Kapral, GJ; Wang, X; Murray, LW; Arendall, WB; Snoeyink, J; Richardson, JS; Richardson, DC
  • How to study proteins by circular dichroism
    Kelly, SM; Jess, TJ; Price, NC
  • Are scoring functions in protein-protein docking ready to predict interactome? Clues from a novel binding affinity benchmark
    Kastritis, PL; Bonvin, AMJJ
  • A key evolutionary mutation enhances DNA binding of the FOXP2 forkhead domain
    Morris, G; Fanucchi, S
  • Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid anteractions
    Hellman, L; Fried, M
  • Stabilizing labile DNA-protein complexes in polyacrylamide gels
    Sidorova, NY; Hung, S; Rau, DC

You’re reading a free preview. Subscribe to read the entire article.

Try 2 weeks free now

DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

or browse the journals available

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off

Sign up
for free
Start 14 day
Free Trial