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Molecular biology of the cell
07 15, 2020;31 (15): 1595-1610.

Flightless anchors IQGAP1 and R-ras to mediate cell extension formation and matrix remodeling.

P D Arora, K Nakajima, A Nanda, A Plaha, A Wilde, D B Sacks, C A McCulloch
Author Information
  1. P D Arora: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
  2. K Nakajima: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
  3. A Nanda: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
  4. A Plaha: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
  5. A Wilde: Departments of Medical Genetics and Biochemistry, Faculty of Medicine, University of Toronto, Toronto, ON M5G 1L7, Canada.
  6. D B Sacks: Department of Laboratory Medicine, National Institutes of Health, Bethesda, MD 20892.
  7. C A McCulloch: Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
PMID: 32432944 DOI: 10.1091/mbc.E19-10-0554

Abstract

Tractional remodeling of collagen fibrils by fibroblasts requires long cell extensions that mediate fibril alignment. The formation of these cell extensions involves flightless I (FliI), an actin-binding protein that contains a leucine-rich-repeat (LRR), which binds R-ras and may regulate cdc42. We considered that FliI interacts with small GTPases and their regulators to mediate assembly of cell extensions. Mass spectrometry analyses of FliI immunoprecipitates showed abundant Ras GTPase-activating-like protein (IQGAP1), which in immunostained samples colocalized with FliI at cell adhesions. Knockdown of IQGAP1 reduced the numbers of cell extensions and the alignment of collagen fibrils. In experiments using dominant negative mutants, cdc42 activity was required for the formation of short extensions while R-ras was required for the formation of long extensions. Immunoprecipitation of wild-type and mutant constructs showed that IQGAP1 associated with cdc42 and R-ras; this association required the GAP-related domain (1004-1237 aa) of IQGAP1. In cells transfected with FliI mutants, the LRR of FliI, but not its gelsolin-like domains, mediated association with cdc42, R-ras, and IQGAP1. We conclude that FliI interacts with IQGAP1 and co-ordinates with cdc42 and R-ras to control the formation of cell extensions that enable collagen tractional remodeling.

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Grants

  1. MOP-36332/CIHR

MeSH Term

3T3 Cells
Animals
Cell Adhesion
Cell Surface Extensions
Collagen
Extracellular Matrix
Mice
Microfilament Proteins
Models, Biological
Protein Binding
Protein Domains
Trans-Activators
cdc42 GTP-Binding Protein
ras GTPase-Activating Proteins
ras Proteins

Chemicals

FlII protein, mouse
IQ motif containing GTPase activating protein 1
Microfilament Proteins
Trans-Activators
ras GTPase-Activating Proteins
Collagen
Rras protein, mouse
cdc42 GTP-Binding Protein
ras Proteins
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 2

Word Cloud

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