Overexpression of (β)-actin is known to alter cell morphology, though its effect on cell motility has not been documented previously. Here we show that overexpressing (β)-actin in myoblasts has striking effects on motility, increasing cell speed to almost double that of control cells. This occurs by increasing the areas of protrusion and retraction and is accompanied by raised levels of (β)-actin in the newly protruded regions. These regions of the cell margin, however, show decreased levels of polymerised actin, indicating that protrusion can outpace the rate of actin polymerisation in these cells. Moreover, the expression of (β)*-actin (a G244D mutant, which shows defective polymerisation in vitro) is equally effective at increasing speed and protrusion. Concomitant changes in actin binding proteins show no evidence of a consistent mechanism for increasing the rate of actin polymerisation in these actin overexpressing cells. The increase in motility is confined to poorly spread cells in both cases and the excess motility can be abolished by blocking myosin function with butanedione monoxime (BDM). Our observations on normal myoblasts are consistent with the view that they protrude by the assembly and cross linking of actin filaments. In contrast, the additional motility shown by cells overexpressing (β)-actin appears not to result from an increase in the rate of actin polymerisation but to depend on myosin function. This suggests that the additional protrusion arises from a different mechanism. We discuss the possibility that it is related to retraction-induced protrusion in fibroblasts. In this phenomenon, a wave of increased protrusion follows a sudden collapse in cell spreading. This view could explain why it is only the additional motility that depends on spreading, and has implications for understanding the differences in locomotion that distinguish tissue cells from highly invasive cell types such as leucocytes and malignant cells.

Brown
A. F.
,
Dunn
G. A.
(
1989
).
Microinterferometry of the movement of dry matter in fibroblasts.
J. Cell Sci
92
,
379
89
Chen
W. T.
(
1979
).
Induction of spreading during fibroblast movement.
J. Cell Biol
81
,
684
691
Chen
W. T.
(
1981
).
Mechanism of retraction of the trailing edge during fibroblast movement.
J. Cell Biol
90
,
187
200
Clark
P.
,
Coles
D.
,
Peckham
M.
(
1997
).
Preferential adhesion to and survival on patterned laminin organizes myogenesis in vitro.
Exp. Cell Res
230
,
275
283
Condeelis
J.
(
1993
).
Life at the leading edge: the formation of cell protrusions.
Annu. Rev. Cell Biol
9
,
411
444
Cramer
L. P.
,
Mitchison
T. J.
(
1995
).
Myosin is involved in postmitotic cell spreading.
J. Cell Biol
131
,
179
89
Cunningham
C. C.
(
1995
).
Actin polymerization and intracellular solvent flow in cell surface blebbing.
J. Cell Biol
129
,
1589
1599
Dunn
G. A.
,
Zicha
D.
(
1995
).
Dynamics of fibroblast spreading.
J. Cell Sci
108
,
1239
1249
Dunn
G. A.
,
Zicha
D.
,
Fraylich
P. E.
(
1997
).
Rapid, microtubule-dependent fluctuations of the cell margin.
J. Cell Sci
110
,
3091
3098
Goldschmidt-Clermont
P. J.
,
Furman
M. I.
,
Wachsstock
D.
,
Safer
D.
,
Nachmias
V. T.
,
Pollard
T. D.
(
1992
).
The control of actin nucleotide exchange by thymosin beta 4 and profilin. A potential regulatory mechanism for actin polymerization in cells.
Mol. Biol. Cell
3
,
1015
1024
Harris
A. K.
(
1973
).
Cell surface movements related to cell locomotion.
Ciba Found. Symp
14
,
3
26
Heacock
C. S.
,
Bamburg
J. R.
(
1983
).
The quantitation of G-and F-actin in cultured cells.
Anal. Biochem
135
,
22
36
Hill
M. A.
,
Gunning
P.
(
1993
).
Beta and gamma actin mRNAs are differentially located within myoblasts.
J. Cell Biol
122
,
825
832
Hoock
T. C.
,
Newcomb
P. M.
,
Herman
I. M.
(
1991
).
Beta actin and its mRNA are localized at the plasma membrane and the regions of moving cytoplasm during the cellular response to injury.
J. Cell Biol
112
,
653
664
Keller
H.
,
Eggli
P.
(
1998
).
Protrusive activity, cytoplasmic compartmentalization, and restriction rings in locomoting blebbing Walker carcinosarcoma cells are related to detachment of cortical actin from the plasma membrane.
Cell Motil. Cytoskel
41
,
181
193
Kislauskis
E. H.
,
Li
Z.
,
Singer
R. H.
,
Taneja
K. L.
(
1993
).
Isoform-specific 3-untranslated sequences sort alpha-cardiac and beta-cytoplasmic actin messenger RNAs to different cytoplasmic compartments.
J. Cell Biol
123
,
165
172
Kislauskis
E. H.
,
Zhu
X.
,
Singer
R. H.
(
1997
).
Beta-actin messenger RNA localization and protein synthesis augment cell motility.
J. Cell Biol
136
,
1263
1270
Laemmli
U. K.
(
1970
).
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
,
680
685
Leavitt
J.
,
Kakunaga
T.
(
1980
).
Expression of a variant form of actin and additional polypeptide changes following chemical-induced in vitro neoplastic transformation of human fibroblasts.
J. Biol. Chem
255
,
1650
1661
Leavitt
J.
,
Gunning
P.
,
Porreca
P.
,
Ng
S. Y.
,
Lin
C. S.
,
Kedes
L.
(
1984
).
Molecular cloning and characterization of mutant and wild-type human beta-actin genes.
Mol. Cell. Biol
4
,
1961
1969
Lloyd
C.
,
Schevzov
G.
,
Gunning
P.
(
1992
).
Transfection of nonmuscle beta-and gamma-actin genes into myoblasts elicits different feedback regulatory responses from endogenous actin genes.
J. Cell Biol
117
,
787
797
Loisel
T. P.
,
Boujemaa
R.
,
Pantaloni
D.
,
Carlier
M. F.
(
1999
).
Reconstitution of actin-based motility of Listeria and Shigella using pure proteins.
Nature
401
,
613
616
Millonig
R.
,
Salvo
H.
,
Aebi
U.
(
1988
).
Probing actin polymerization by intermolecular cross-linking.
J. Cell Biol
106
,
785
796
Minamide
L. S.
,
Painter
W. B.
,
Schevzov
G.
,
Gunning
P.
,
Bamburg
J. R.
(
1997
).
Differential regulation of actin depolymerizing factor and cofilin in response to alterations in the actin monomer pool.
J. Biol. Chem
272
,
8303
8309
Mitchison
T. J.
,
Cramer
L. P.
(
1996
).
Actin-based cell motility and cell locomotion.
Cell
84
,
371
379
Mogilner
A.
,
Oster
G.
(
1996
).
Cell motility driven by actin polymerization.
Biophys. J
71
,
3030
3045
Morgan
J. E.
,
Beauchamp
J. R.
,
Pagel
C. N.
,
Peckham
M.
,
Ataliotis
P.
,
Jat
P. S.
,
Noble
M. D.
,
Farmer
K.
,
Partridge
T. A.
(
1994
).
Myogenic cell lines derived from transgenic mice carrying a thermolabile T antigen: a model system for the derivation of tissue-specific and mutation-specific cell lines.
Dev. Biol
162
,
486
498
Schagger
H.
,
von Jagow
G.
(
1987
).
Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
Anal. Biochem
166
,
368
379
Schevzov
G.
,
Lloyd
C.
,
Gunning
P.
(
1992
).
High level expression oftransfected beta-and gamma-actin genes differentially impacts on myoblast cytoarchitecture.
J. Cell Biol
117
,
775
785
Small
J. V.
(
1981
).
Organization of actin in the leading edge of culture cells: influence of osmium tetroxide and dehydration on the ultrastructure of actin meshworks.
J. Cell Biol
91
,
695
705
Small
J. V.
,
Rohlfs
A.
,
Herzog
M.
(
1993
).
Actin and cell movement.
Symp. Soc. Exp. Biol
47
,
57
71
Stossel
T. P.
,
Hartwig
J. H.
,
Janmey
P. A.
,
Kwiatkowski
D. J.
(
1999
).
Cell crawling two decades after Abercrombie.
Biochem. Soc. Symp
65
,
267
280
Svitkina
T. M.
,
Neyfakh
A. A.
Jr.
,
Bershadsky
A. D.
(
1986
).
Actin cytoskeleton of spread fibroblasts appears to assemble at the cell edges.
J. Cell Sci
82
,
235
248
Taniguchi
S.
,
Sagara
J.
,
Kakunaga
T.
(
1988
).
Deficient polymerization in vitro of a point-mutated beta-actin expressed in a transformed human fibroblast cell line.
J. Biochem. (Tokyo)
103
,
707
713
Wang
E.
(
1985
).
Are cross-bridging structures involved in the bundle formation of intermediate filaments and the decrease in locomotion that accompany cell aging?.
J. Cell Biol
100
,
1466
1473
Wells
C.
,
Coles
D.
,
Entwistle
A.
,
Peckham
M.
(
1997
).
Myogenic cells express multiple myosin isoforms.
J. Muscle Res. Cell Motil
18
,
501
515
Zicha
D.
,
Dunn
G. A.
(
1995
).
An image processing system for cell behaviour studies in subconfluent cultures.
J. Micros
179
,
11
21
This content is only available via PDF.