Interaction with the IQ3 motif of myosin-10 is required for calmodulin-like protein-dependent filopodial extension

Richard D. Bennett; Ariel J. Caride; Amy S. Mauer; Emanuel E. Strehler

doi:10.1016/j.febslet.2008.05.051

Interaction with the IQ3 motif of myosin-10 is required for calmodulin-like protein-dependent filopodial extension

Richard D. Bennett, Ariel J. Caride, Amy S. Mauer, Emanuel E. Strehler

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

Calmodulin-like protein (CLP) is a specific light chain of unconventional myosin-10 (Myo10) and enhances Myo10-dependent filopodial extension. Here we show that phenylalanine-795 in the third IQ domain (IQ3) of Myo10 is critical for CLP binding. Remarkably, mutation of F795 to alanine had little effect on calmodulin binding to IQ3. Fluorescence microscopy and time-lapse video microscopy showed that HeLa cells expressing CLP and transiently transfected with GFP-Myo10-F795A exhibited significantly shorter filopodia and decreased intrafilopodial motility compared to wildtype GFP-Myo10-transfected cells. Thus, F795 represents a unique anchor for CLP and is essential for CLP-mediated Myo10 function in filopodial extension and motility. Structured summary: MINT-6595901: GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CLP (uniprotkb:P27482) by pull down (MI:0096)MINT-6596000: CLP (uniprotkb:P27482) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6596013: CaM (uniprotkb:P62158) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6595938:GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CaM (uniprotkb:P62158) by pull down (MI:0096).

Original language	English (US)
Pages (from-to)	2377-2381
Number of pages	5
Journal	FEBS Letters
Volume	582
Issue number	16
DOIs	https://doi.org/10.1016/j.febslet.2008.05.051
State	Published - Jul 9 2008

Keywords

Calmodulin-like protein
Filopodia
IQ domain
Motility
Myosin-10

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Access to Document

10.1016/j.febslet.2008.05.051

Cite this

@article{a221ff156186460e90a117f4918226d2,

title = "Interaction with the IQ3 motif of myosin-10 is required for calmodulin-like protein-dependent filopodial extension",

abstract = "Calmodulin-like protein (CLP) is a specific light chain of unconventional myosin-10 (Myo10) and enhances Myo10-dependent filopodial extension. Here we show that phenylalanine-795 in the third IQ domain (IQ3) of Myo10 is critical for CLP binding. Remarkably, mutation of F795 to alanine had little effect on calmodulin binding to IQ3. Fluorescence microscopy and time-lapse video microscopy showed that HeLa cells expressing CLP and transiently transfected with GFP-Myo10-F795A exhibited significantly shorter filopodia and decreased intrafilopodial motility compared to wildtype GFP-Myo10-transfected cells. Thus, F795 represents a unique anchor for CLP and is essential for CLP-mediated Myo10 function in filopodial extension and motility. Structured summary: MINT-6595901: GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CLP (uniprotkb:P27482) by pull down (MI:0096)MINT-6596000: CLP (uniprotkb:P27482) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6596013: CaM (uniprotkb:P62158) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6595938:GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CaM (uniprotkb:P62158) by pull down (MI:0096).",

keywords = "Calmodulin-like protein, Filopodia, IQ domain, Motility, Myosin-10",

author = "Bennett, {Richard D.} and Caride, {Ariel J.} and Mauer, {Amy S.} and Strehler, {Emanuel E.}",

note = "Funding Information: We thank Dr. Katalin T{\"o}r{\"o}k (St. Luke{\textquoteright}s Children Hospital, London, UK) for the generous gift of TA-CaM and TA-CLP. This work was supported in part by a grant from the Susan G. Komen Breast Cancer Foundation (BCTR-0201979) and by the Mayo Graduate School. ",

year = "2008",

month = jul,

day = "9",

doi = "10.1016/j.febslet.2008.05.051",

language = "English (US)",

volume = "582",

pages = "2377--2381",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Elsevier",

number = "16",

}

TY - JOUR

T1 - Interaction with the IQ3 motif of myosin-10 is required for calmodulin-like protein-dependent filopodial extension

AU - Bennett, Richard D.

AU - Caride, Ariel J.

AU - Mauer, Amy S.

AU - Strehler, Emanuel E.

N1 - Funding Information: We thank Dr. Katalin Török (St. Luke’s Children Hospital, London, UK) for the generous gift of TA-CaM and TA-CLP. This work was supported in part by a grant from the Susan G. Komen Breast Cancer Foundation (BCTR-0201979) and by the Mayo Graduate School.

PY - 2008/7/9

Y1 - 2008/7/9

N2 - Calmodulin-like protein (CLP) is a specific light chain of unconventional myosin-10 (Myo10) and enhances Myo10-dependent filopodial extension. Here we show that phenylalanine-795 in the third IQ domain (IQ3) of Myo10 is critical for CLP binding. Remarkably, mutation of F795 to alanine had little effect on calmodulin binding to IQ3. Fluorescence microscopy and time-lapse video microscopy showed that HeLa cells expressing CLP and transiently transfected with GFP-Myo10-F795A exhibited significantly shorter filopodia and decreased intrafilopodial motility compared to wildtype GFP-Myo10-transfected cells. Thus, F795 represents a unique anchor for CLP and is essential for CLP-mediated Myo10 function in filopodial extension and motility. Structured summary: MINT-6595901: GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CLP (uniprotkb:P27482) by pull down (MI:0096)MINT-6596000: CLP (uniprotkb:P27482) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6596013: CaM (uniprotkb:P62158) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6595938:GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CaM (uniprotkb:P62158) by pull down (MI:0096).

AB - Calmodulin-like protein (CLP) is a specific light chain of unconventional myosin-10 (Myo10) and enhances Myo10-dependent filopodial extension. Here we show that phenylalanine-795 in the third IQ domain (IQ3) of Myo10 is critical for CLP binding. Remarkably, mutation of F795 to alanine had little effect on calmodulin binding to IQ3. Fluorescence microscopy and time-lapse video microscopy showed that HeLa cells expressing CLP and transiently transfected with GFP-Myo10-F795A exhibited significantly shorter filopodia and decreased intrafilopodial motility compared to wildtype GFP-Myo10-transfected cells. Thus, F795 represents a unique anchor for CLP and is essential for CLP-mediated Myo10 function in filopodial extension and motility. Structured summary: MINT-6595901: GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CLP (uniprotkb:P27482) by pull down (MI:0096)MINT-6596000: CLP (uniprotkb:P27482) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6596013: CaM (uniprotkb:P62158) and GST-IQ3 Myo10 (uniprotkb:Q9HD67) bind (MI:0407) by classical fluorescence spectroscopy (MI:0017)MINT-6595938:GST-IQ3 Myo10 (uniprotkb:Q9HD67) binds (MI:0407) to CaM (uniprotkb:P62158) by pull down (MI:0096).

KW - Calmodulin-like protein

KW - Filopodia

KW - IQ domain

KW - Motility

KW - Myosin-10

UR - http://www.scopus.com/inward/record.url?scp=45549109551&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=45549109551&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2008.05.051

DO - 10.1016/j.febslet.2008.05.051

M3 - Article

C2 - 18570893

AN - SCOPUS:45549109551

SN - 0014-5793

VL - 582

SP - 2377

EP - 2381

JO - FEBS Letters

JF - FEBS Letters

IS - 16

ER -

Interaction with the IQ3 motif of myosin-10 is required for calmodulin-like protein-dependent filopodial extension

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this