Identification of FHOD1-binding proteins and mechanisms of FHOD1-regulated actin dynamics

Jennifer J Westendorf, Sreenivas Koka

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Formin homology-2-domain containing protein 1 (FHOD1) regulates gene transcription, actin-cytoskeleton structure, and cell migration. To gain insight into the mechanisms by which FHOD1 mediates these diverse activities, a yeast-two-hybrid screen was performed to identify FHOD1-binding proteins. Three proteins specifically interacted with the carboxy-terminal two-thirds of FHOD1, which includes the FH1, FH2, and diaphanous activating domains (DAD). The newly identified FHOD1-binding proteins are protein kinase C binding protein 1 (PRKCBP1), cyclophilin B, and an isoform of WASP-interacting SH3-domain protein/diaphanous-interacting protein 1 (WISH/DIP1), named WISH-B. The proline-rich FH1 domain of FHOD1 was sufficient to interact with the central portion of PRKCP1 and full-length cyclophilin B. The FH1 domain also interacted with full-length WISH-B, but the extreme amino-terminus was sufficient to associate with WISH-B as well. WISH-B altered the solubility of FHOD1 in vitro and a truncation mutant containing the amino-terminal 227 residues of WISH-B disrupted FHOD1-induced stress fibers. WISH-B did not affect FHOD1-induced gene transcription through the serum response factor (SRF) recognition site on the skeletal α-actin promoter (SkA). However, stabilization of F-actin prevented FHOD1 dependent activation of this promoter in presence of high, but not low serum concentrations. Thus, the identification of a new FHOD1-binding protein provides insight into the mechanisms by which FHOD1 regulates actin polymerization and transcription.

Original languageEnglish (US)
Pages (from-to)29-41
Number of pages13
JournalJournal of Cellular Biochemistry
Volume92
Issue number1
DOIs
StatePublished - 2004
Externally publishedYes

Fingerprint

Protein Binding
Actins
Carrier Proteins
Proteins
Transcription
Serum Response Factor
Protein Domains
Stress Fibers
src Homology Domains
Actin Cytoskeleton
Proline
Polymerization
Solubility
Protein Kinase C
Genes
Cell Movement
Protein Isoforms
Yeasts
Yeast
Serum

Keywords

  • Cyclophilin B
  • Diaphanous
  • DIP1
  • FHOD1
  • FHOS
  • Formin
  • PRKCP1
  • RACK7
  • Stress fibers
  • WISH

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Identification of FHOD1-binding proteins and mechanisms of FHOD1-regulated actin dynamics. / Westendorf, Jennifer J; Koka, Sreenivas.

In: Journal of Cellular Biochemistry, Vol. 92, No. 1, 2004, p. 29-41.

Research output: Contribution to journalArticle

@article{1692e0ac64de439890ab3bfda741856f,
title = "Identification of FHOD1-binding proteins and mechanisms of FHOD1-regulated actin dynamics",
abstract = "Formin homology-2-domain containing protein 1 (FHOD1) regulates gene transcription, actin-cytoskeleton structure, and cell migration. To gain insight into the mechanisms by which FHOD1 mediates these diverse activities, a yeast-two-hybrid screen was performed to identify FHOD1-binding proteins. Three proteins specifically interacted with the carboxy-terminal two-thirds of FHOD1, which includes the FH1, FH2, and diaphanous activating domains (DAD). The newly identified FHOD1-binding proteins are protein kinase C binding protein 1 (PRKCBP1), cyclophilin B, and an isoform of WASP-interacting SH3-domain protein/diaphanous-interacting protein 1 (WISH/DIP1), named WISH-B. The proline-rich FH1 domain of FHOD1 was sufficient to interact with the central portion of PRKCP1 and full-length cyclophilin B. The FH1 domain also interacted with full-length WISH-B, but the extreme amino-terminus was sufficient to associate with WISH-B as well. WISH-B altered the solubility of FHOD1 in vitro and a truncation mutant containing the amino-terminal 227 residues of WISH-B disrupted FHOD1-induced stress fibers. WISH-B did not affect FHOD1-induced gene transcription through the serum response factor (SRF) recognition site on the skeletal α-actin promoter (SkA). However, stabilization of F-actin prevented FHOD1 dependent activation of this promoter in presence of high, but not low serum concentrations. Thus, the identification of a new FHOD1-binding protein provides insight into the mechanisms by which FHOD1 regulates actin polymerization and transcription.",
keywords = "Cyclophilin B, Diaphanous, DIP1, FHOD1, FHOS, Formin, PRKCP1, RACK7, Stress fibers, WISH",
author = "Westendorf, {Jennifer J} and Sreenivas Koka",
year = "2004",
doi = "10.1002/jcb.20031",
language = "English (US)",
volume = "92",
pages = "29--41",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "1",

}

TY - JOUR

T1 - Identification of FHOD1-binding proteins and mechanisms of FHOD1-regulated actin dynamics

AU - Westendorf, Jennifer J

AU - Koka, Sreenivas

PY - 2004

Y1 - 2004

N2 - Formin homology-2-domain containing protein 1 (FHOD1) regulates gene transcription, actin-cytoskeleton structure, and cell migration. To gain insight into the mechanisms by which FHOD1 mediates these diverse activities, a yeast-two-hybrid screen was performed to identify FHOD1-binding proteins. Three proteins specifically interacted with the carboxy-terminal two-thirds of FHOD1, which includes the FH1, FH2, and diaphanous activating domains (DAD). The newly identified FHOD1-binding proteins are protein kinase C binding protein 1 (PRKCBP1), cyclophilin B, and an isoform of WASP-interacting SH3-domain protein/diaphanous-interacting protein 1 (WISH/DIP1), named WISH-B. The proline-rich FH1 domain of FHOD1 was sufficient to interact with the central portion of PRKCP1 and full-length cyclophilin B. The FH1 domain also interacted with full-length WISH-B, but the extreme amino-terminus was sufficient to associate with WISH-B as well. WISH-B altered the solubility of FHOD1 in vitro and a truncation mutant containing the amino-terminal 227 residues of WISH-B disrupted FHOD1-induced stress fibers. WISH-B did not affect FHOD1-induced gene transcription through the serum response factor (SRF) recognition site on the skeletal α-actin promoter (SkA). However, stabilization of F-actin prevented FHOD1 dependent activation of this promoter in presence of high, but not low serum concentrations. Thus, the identification of a new FHOD1-binding protein provides insight into the mechanisms by which FHOD1 regulates actin polymerization and transcription.

AB - Formin homology-2-domain containing protein 1 (FHOD1) regulates gene transcription, actin-cytoskeleton structure, and cell migration. To gain insight into the mechanisms by which FHOD1 mediates these diverse activities, a yeast-two-hybrid screen was performed to identify FHOD1-binding proteins. Three proteins specifically interacted with the carboxy-terminal two-thirds of FHOD1, which includes the FH1, FH2, and diaphanous activating domains (DAD). The newly identified FHOD1-binding proteins are protein kinase C binding protein 1 (PRKCBP1), cyclophilin B, and an isoform of WASP-interacting SH3-domain protein/diaphanous-interacting protein 1 (WISH/DIP1), named WISH-B. The proline-rich FH1 domain of FHOD1 was sufficient to interact with the central portion of PRKCP1 and full-length cyclophilin B. The FH1 domain also interacted with full-length WISH-B, but the extreme amino-terminus was sufficient to associate with WISH-B as well. WISH-B altered the solubility of FHOD1 in vitro and a truncation mutant containing the amino-terminal 227 residues of WISH-B disrupted FHOD1-induced stress fibers. WISH-B did not affect FHOD1-induced gene transcription through the serum response factor (SRF) recognition site on the skeletal α-actin promoter (SkA). However, stabilization of F-actin prevented FHOD1 dependent activation of this promoter in presence of high, but not low serum concentrations. Thus, the identification of a new FHOD1-binding protein provides insight into the mechanisms by which FHOD1 regulates actin polymerization and transcription.

KW - Cyclophilin B

KW - Diaphanous

KW - DIP1

KW - FHOD1

KW - FHOS

KW - Formin

KW - PRKCP1

KW - RACK7

KW - Stress fibers

KW - WISH

UR - http://www.scopus.com/inward/record.url?scp=16244379726&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=16244379726&partnerID=8YFLogxK

U2 - 10.1002/jcb.20031

DO - 10.1002/jcb.20031

M3 - Article

C2 - 15095401

AN - SCOPUS:16244379726

VL - 92

SP - 29

EP - 41

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 1

ER -