The rat homologue of Wiskott-Aldrich syndrome protein (WASP)-interacting protein (WIP) associates with actin filaments, recruits N-WASP from the nucleus, and mediates mobilization of actin from stress fibers in favor of filopodia formation.
We cloned and characterized the rat homologue of the Wiskott-Aldrich syndrome protein (WASP)-interacting protein (WIP). Rat WIP shows 86% amino acid sequence identity to human WIP. Northern analyses revealed two major mRNA species of 5.0 and 3.8 kb, which were ubiquitously expressed, though predominantly in spleen and lung. Minor species of 2.4, 1.8, 1.4, and 1.1 kb were also detected in some tissues and cell lines. Thus, WIP is subject to tissue-specific alternative splicing. WIP bound to N-WASP in vivo, as revealed by co-immunoprecipitation. Expression of WIP in rat fibroblasts revealed a clear co-localization with actin stress fibers. However, expression in tumor cells lacking actin cables did not restore these structures. Interestingly, co-expression of WIP and N-WASP resulted in redistribution of N-WASP, abrogating its dominant nuclear expression and leading to co-localization with WIP in the perinuclear area and with actin in membrane protrusions. Moreover, stress fibers and, concomitantly, the associated WIP were largely dissolved. Very similar effects were seen upon epidermal growth factor stimulation of serum-starved cells. Our results suggest that WIP might be involved in transmitting mitogenic signals to cytoskeletal functions, perhaps by modulating the subcellular localization of N-WASP. Interaction of N-WASP with WIP may in turn lead to mobilization of actin from stress fibers and nucleation of new actin filaments in filopodia.
Vetterkind
Susanne
S
Institute of Genetics, University of Bonn, Roemerstrasse 164, D-53117 Bonn, Germany.
Miki
Hiroaki
H
Takenawa
Tadaomi
T
Klawitz
Ingrid
I
Scheidtmann
Karl-Heinz
KH
Preuss
Ute
U
D000199:Actins / Q000378:metabolism*; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism*; D011554:Pseudopodia / Q000502:physiology*; D051381:Rats; D022502:Stress Fibers / Q000378:metabolism*; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal
D000199:Actins; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; C495771:WASL protein, human; C109903:WIPF1 protein, human; C495770:Wasl protein, rat; C491670:Wipf1 protein, rat; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D004815:Epidermal Growth Factor