We used immunological approaches to study the factors controlling the distribution of the Na,K-ATPase in fast twitch skeletal muscle of the rat. Both alpha subunits of the Na,K-ATPase colocalize with beta-spectrin and ankyrin 3 in costameres, structures at the sarcolemma that lie over Z and M-lines and in longitudinal strands. In immunoprecipitates, the alpha1 and alpha2 subunits of the Na,K-ATPase as well as ankyrin 3 associate with beta-spectrin/alpha- fodrin heteromers and with a pool of beta-spectrin at the sarcolemma that does not contain alpha-fodrin. Myofibers of mutant mice lacking beta-spectrin (ja/ja) have a more uniform distribution of both the alpha1 and alpha2 subunits of the Na,K-ATPase in the sarcolemma, supporting the idea that the rectilinear sarcomeric pattern assumed by the Na,K-ATPase in wild-type muscle requires beta-spectrin. The Na,K-ATPase and beta-spectrin are distributed normally in muscle fibers of the nb/nb mouse, which lacks ankyrin 1, suggesting that this isoform of ankyrin is not necessary to link the Na,K-ATPase to the spectrin-based membrane skeleton. In immunofluorescence and subcellular fractionation experiments, the alpha2 but not the alpha1 subunit of the Na,K-ATPase is present in transverse (t-) tubules. The alpha1 subunit of the pump is not detected in increased amounts in the t-tubules of muscle from the ja/ja mouse, however. Our results suggest that the spectrin-based membrane skeleton, including ankyrin 3, concentrates both isoforms of the Na,K-ATPase in costameres, but that it does not play a significant role in restricting the entry of the alpha1 subunit into the t-tubules.
REFERENCES
Bennett
V.
(1990
). Spectrin-based membrane skeleton, a multipotential adaptor between plasma membrane and cytoplasm.
Physiol. Rev.
70
, 1029
–1065
Birkenmeier
C. S.
, White
R. A.
, Peters
L. L.
, Hall
E. J.
, Lux
S. E.
, Barker
J. E.
(1993
). Complex patterns of sequence variation and multiple5 and 3ends are found among transcripts of the erythroid ankyrin gene.
J. Biol. Chem
268
, 9533
–9540
Birkenmeier
C. S.
, Sharp
J. J.
, Gifford
E. J.
, Deveau
S. A.
, Barker
J. E.
(1998
). An alternative first exon in the distal end of the erythroid ankyrin gene leads to production of a small isoform containing an NH2-terminal membrane anchor.
Genomics
50
, 79
–88
Bloom
M. L.
, Kaysser
T. M.
, Birkenmeier
C. S.
, Barker
J. E.
(1994
). The murine mutation jaundiced is caused by replacement of an arginine with a stop codon in the mRNA encoding the ninth repeat of beta-spectrin.
Proc. Nat. Acad. Sci. USA
91
, 10099
–10103
Bodine
D. M.
, Birkenmeier
C. S.
, Barker
J. E.
(1984
). Spectrin deficient inherited hemolytic anemias in the mouse, Characterization by spectrin synthesis and mRNA activity in reticulocytes.
Cell
37
, 721
–729
Burnette
W. N.
(1981
). ‘Western blotting’, Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated Protein A.
Anal. Biochem
112
, 195
–203
Colognato
H.
, Winkelmann
D. A.
, Yurchenco
P. D.
(1999
). Laminin polymerization induces a receptor-cytoskeleton network.
J. Cell Biol
145
, 619
–631
Craig
S. W.
, Pardo
J. V.
(1983
). Gamma actin, spectrin, and intermediate filament proteins colocalize with vinculin at costameres, myofibril-to-sarcolemma attachment sites.
Cell Motil
3
, 449
–462
Davis
J. Q.
, Bennett
V.
(1990
). The anion exchanger and Na+,K+-ATPase interact with distinct sites on ankyrin in in vitro assays.
J. Biol. Chem
265
, 17252
–17256
Davis
J. Q.
, Bennett
V.
(1993
). Ankyrin-binding activity of nervous system cell adhesion molecules expressed in adult brain.
J. Cell Sci. Suppl
17
, 109
–117
Devarajan
P.
, Scaramuzzino
D. A.
, Morrow
J. S.
(1994
). Ankyrin binds to two distinct cytoplasmic domains of Na, K-ATPase alpha subunit.
Proc. Nat. Acad. Sci. USA
91
, 2965
–2969
Devarajan
P.
, Stabach
P. R.
, Mann
A. S.
, Ardito
T.
, Kashgarian
M.
, Morrow
J. S.
(1996
). Identification of a small cytoplasmic ankyrin (AnkG119) in the kidney and muscle that bindsI * spectrin and associates with the Golgi apparatus.
J. Cell Biol
133
, 819
–830
Ehmer
S.
, Herrmann
R.
, Bittner
R.
, Voit
T.
(1997
). Spatial distribution of-spectrin in normal and dystrophic human skeletal musle.
Acta Neuropathol
94
, 240
–246
Felsenfeld
D. P.
, Sweadner
K. J.
(1998
). Fine specificity mapping and topography of an isozyme-specific epitope of the Na, K-ATPase catalytic subunit.
J. Biol. Chem
263
, 10932
–10942
Flucher
B. E.
, Daniels
M. P.
(1989
). Distribution of Na+channel and ankyrin in neuromuscular junction is complementary to acetylcholine receptor and 43-kD protein.
Neuron
3
, 163
–175
Gallagher
P. G.
, Forget
B. G.
(1993
). Spectrin genes in health and disease.
Semin. Hematol
30
, 4
–21
Georgatos
S. D.
, Blobel
G.
(1987
). Two distinct attachment sites for vimentin along the plasma membrane and the nuclear envelope in avain erythroyctes: A basis for a vectorial assembly of intermediate filaments.
J. Cell Biol
105
, 105
–115
Granger
B. L.
, Lazarides
E.
(1979
). Desmin and vimentin coexist at the periphery of the myofibril Z disc.
Cell
18
, 1053
–1063
Hartwig
J. H.
(1995
). Actin-binding proteins. 1: Spectrin super family.
Prot. Profile
2
, 703
–800
Hassoun
H.
, Palek
J.
(1996
). Hereditary spherocytosis, a review of the clinical and molecular aspects of the disease.
Blood Rev.
10
, 129
–47
Hoffman
E. P.
, Brown
R. H.
Jr., Kunkel
L. M.
(1987
). Dystrophin, The protein product of the Duchenne Muscular Dystrophy locus.
Cell
51
, 919
–928
Hoffman
E. P.
, Watkins
S. C.
, Slayter
H. S.
, Kunkel
L. M.
(1989
). Detection of a specific isoform of-actinin with antisera directed against dystrophin.
J. Cell Biol
108
, 503
–510
Hu
R.-J.
, Moorthy
S.
, Bennett
V.
(1995
). Expression of functional domains ofG-spectrin disrupts epithelial morphology in cultured cells.
J. Cell Biol
128
, 1069
–1080
Jordan
C.
, Puschel
B.
, Koob
R.
, Drenckhahn
D.
(1995
). Identification of a binding motif for ankyrin on the alpha-subunit of Na+,K+-ATPase.
J. Biol. Chem
270
, 29971
–29975
Kaysser
T. M.
, Wandersee
N. J.
, Bronson
R. T.
, Barker
J. E.
(1997
). Thrombosis and secondary hemochromatosis play major roles in the pathogenesis of jaundiced and spherocytic mice, murine models for hereditary spherocytosis.
Blood
90
, 4610
–4619
Kennedy
S. P.
, Warren
S. L.
, Forget
B. L.
, Morrow
J. S.
(1991
). Ankyrin binds to the 15th repetitive unit of erythroid and nonerythroid beta-spectrin.
J. Cell Biol
115
, 267
–277
Koob
R.
, Kraemer
D.
, Trippe
G.
, Aebi
U.
, Drenckhahn
D.
(1990
). Association of kidney and parotid Na+, K+-ATPase microsomes with actin and analogs of spectrin and ankyrin.
Eur. J. Cell Biol
53
, 93
–100
Kordeli
E.
, Ludosky
M. A.
, Deprette
C.
, Frappier
T.
, Cartaud
J.
(1998
). AnkyrinG is associated with the postsynaptic membrane and the sarcoplasmic reticulum in the skeletal muscle fiber.
J. Cell Sci
111
, 2197
–207
Laemmli
U. K.
(1970
). Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
, 680
–685
Langley
R. C.
Jr., Cohen
C. M.
(1986
). Association of spectrin with desmin intermediate filaments.
J. Cell Biochem
30
, 101
–109
Langley
R. C.
Jr., Cohen
C. M.
(1987
). Cell type-specific association between two types of spectrin and two types of intermediate filaments.
Cell Motil. Cytoskel
8
, 165
–173
Lavoie
L.
, Roy
D.
, Ramlal
T.
, Dombrowski
L.
, Martin-Vassallo
P.
, Marette
A.
, Carpentier
J. L.
, Klip
A.
(1996
). Insulin-induced translocation of Na+-K+-ATPase subunits to the plasma membrane is muscle fiber type specific.
Am. J. Physiol
270
, 1421
–.Lavoie
L.
, Levenson
R.
, Martin-Vassallo
P.
, Klip
A.
(1997
). The molar ratios of alpha and beta subunits of the Na+-K+-ATPase differ in distinct subcellular membranes from rat skeletal muscle.
Biochemistry
36
, 7726
–7732
Lazarides
E.
(1978
). The distribution of desmin (100 D) filaments in primary cultures of embryonic chick cardiac cells.
Exp. Cell Res
112
, 265
–273
Li
Z.
, Burke
E. P.
, Frank
J. S.
, Bennett
V.
, Philipson
K. D.
(1993
). The cardiac Na+-Ca2+exchanger binds to the cytoskeletal protein ankyrin.
J. Biol. Chem
268
, 11489
–11491
Madreperla
S. A.
, Edidin
M.
, Adler
R.
(1989
). Na+,K+-adenosine triphosphatase polarity in retinal photoreceptors: a role for cytoskeletal attachments.
J. Cell Biol
109
, 1483
–1493
Morrow
J. S.
, Cianci
C. D.
, Ardito
T.
, Mann
A. S.
, Kashgarian
M.
(1989
). Ankyrin links fodrin to the alpha subunit of Na, K-ATPase in Madin-Darby canine kidney cells and intact renal tubule cells.
J. Cell Biol
108
, 455
–465
Nelson
W. J.
, Lazarides
E.
(1983
). Expression of thesubunit of spectrin in nonerythroid cells.
Proc. Nat. Acad. Sci. USA
80
, 363
–367
Nelson
W. J.
, Lazarides
E.
(1984
). Goblin (ankyrin) in striated muscle: identification of the potential membrane receptor for erythroid spectrin in muscle cells.
Proc. Nat. Acad. Sci. USA
81
, 3292
–3296
Nelson
W. J.
, Veshnock
P. J.
(1987
). Ankyrin binding to (Na++K+) ATPase and implications for the organization of membrane domains in polarized cells.
Nature
328
, 533
–536
Nelson
W. J.
, Hammerton
R. W.
(1989
). A membrane-cytoskeletal complex containing Na+,K+-ATPase, ankyrin, and fodrin in Madin-Darby canine kidney (MDCK) cells, implications for the biogenesis of epithelial cell polarity.
J. Cell Biol
108
, 893
–902
Orlowski
J.
, Lingrel
J. B.
(1988
). Differential expression of the Na,K-ATPase alpha 1 and alpha 2 subunit genes in a murine myogenic cell line. Induction of the alpha 2 isozyme during myocyte differentiation.
J. Biol. Chem
263
, 17817
–21
Pardo
J. V.
, Siliciano
J. D.
, Craig
S. W.
(1983
). A vinculin-containing cortical lattice in skeletal muscle: Transverse lattice elements (‘costameres’) mark sites of attachment between myofibrils and sarcolemma.
Proc. Nat. Acad. Sci. USA
80
, 1008
–10012
Peters
L. L.
, John
K. M.
, Lu
F. M.
, Eicher
E. M.
, Higgins
A.
, Yialamas
M.
, Turtzo
L. C.
, Otsuka
A. J.
, Lux
S. E.
(1995
). Ank3 (epithelial ankyrin), a widely distributed new member of the ankyrin gene family and the major ankyrin in kidney, is expressed in alternatively spliced forms, including forms that lack the repeat domain.
J. Cell Biol
130
, 313
–330
Pierobon-Bormioli
S.
(1981
). Transverse sarcomere filamentous systems: ‘Z-and M-cables’.
J. Musc. Res. Cell Motil
2
, 401
–413
Porter
G. A.
, Dmytrenko
G. M.
, Winkelmann
J. C.
, Bloch
R. J.
(1992
). Dystrophin colocalizes with-spectrin in distinct subsarcolemmal domains in mammalian skeletal muscle.
J. Cell Biol
117
, 997
–1005
Porter
G. A.
, Scher
M. G.
, Resneck
W. G.
, Fowler
V.
, Bloch
R. J.
(1997
). Two populations of-spectrin in mammalian skeletal muscle.
Cell Motil. Cytoskel
37
, 7
–19
Richardson
G. P.
, Stromer
M. H.
, Huiatt
T. W.
, Robson
R. M.
(1981
). Immunoelectron and immunofluorescence localization of desmin in mature avian muscles.
Eur. J. Cell Biol
26
, 91
–101
Sands
M. S.
, Barker
J. E.
(1999
). Percutaneous intravenous injection in neonatal mice.
Lab. Anim. Sci
49
, 328
–330
Shear
C. R.
, Bloch
R. J.
(1985
). Vinculin in subsarcolemmal densities in chicken skeletal muscle: localization and relationship to intracellular and extracellular structures.
J. Cell Biol
101
, 240
–256
Srinivasan
Y.
, Elmer
L.
, Davis
J.
, Bennett
V.
, Angelides
K.
(1988
). Ankyrin and spectrin associate with voltage-dependent sodium channels in brain.
Nature
333
, 177
–180
Street
S. F.
(1983
). Lateral transmission of tension in frog myofibers: a myofibrillar network and transverse cytoskeletal connections are possible transmitters.
J. Cell. Physiol
114
, 346
–364
Thevananther
S.
, Kolli
A. H.
, Devarajan
P.
(1998
). Identification of a novel ankyrin isoform (AnkG190) in kidney and lung that associates with the plasma membrane and binds-Na, K-ATPase.
J. Biol. Chem
273
, 23952
–23958
Tsakiridis
T.
, Wong
P. P.
, Liu
Z.
, Rodgers
C.D.
, Vranic
M.
, Klip
A.
(1996
). Exercise increases the plasma membrane content of the Na+-K+-ATPase and its mRNA in rat skeletal muscles.
J. Appl. Physiol
80
, 699
–705
Tuvia
S.
, Buhusi
M.
, Davis
L.
, Reedy
M.
, Bennett
V.
(1999
). Ankyrin-B is required for intracellular sorting of structurally diverse Ca2+homeostasis proteins.
J. Cell Biol
147
, 995
–1008
Urayama
O.
, Shutt
H.
, Sweadner
K. J.
(1989
). Identification of three isozyme proteins of the catalytic subunit of the Na, K-ATPase in rat brain.
J. Biol. Chem
264
, 8271
–80
Viel
A.
, Branton
D.
(1996
). Spectrin: on the path from structure to function.
Curr. Opin. Cell Biol
8
, 49
–55
Vybiral
T.
, Winkelmann
J. C.
, Roberts
R.
, Joe
E.-H.
, Casey
D. L.
, Williams
J. K.
, Epstein
H. F.
(1992
). Human cardiac and skeletal muscle spectrins: differential expression and localization.
Cell Motil. Cytoskel
21
, 293
–304
White
R. A.
, Birkenmeier
C. S.
, Lux
S. E.
, Barker
J. E.
(1990
). Ankyrin and the hemolytic anemia mutation, nb, map to mouse chromosome 8, presence of the nb allele is associated with a truncated erythrocyte ankyrin.
Proc. Nat. Acad. Sci. USA
87
, 3117
–3121
Williams
M. W.
, Bloch
R. J.
(1999
). Extensive but coordinated reorganization of the membrane skeleton in myofibers of dystrophic (mdx) mice.
J. Cell Biol
144
, 1259
–1270
Williams
M. W.
, Bloch
R. J.
(1999
). Differential distribution of dystrophin and-spectrin at the sarcolemma of fast twitch skeletal muscle fibers.
J. Musc. Res. Cell Motil
20
, 387
–398
Williams
M. W.
, Resneck
W. G.
, Bloch
R. J.
(2000
). The membrane skeleton of innervated and denervated fast-and slow-twitch muscle.
Muscle & Nerve
23
, 590
–599
Winkelmann
J. C.
, Costa
F. F.
, Linzie
B. L.
, Forget
B. G.
(1990
). Beta-spectrin in human skeletal muscle. Tissue-specific differential processing of a 3beta-spectrin pre-mRNA generates a beta-spectrin isoform with a unique carboxyl terminus.
J. Biol. Chem
265
, 20449
–20454
Winkelmann
J. C.
, Forget
B. G.
(1993
). Erythroid and nonerythroid spectrins.
Blood
81
, 3173
–3185
Wood
S. J.
, Slater
C. R.
(1998
). -Spectrin is colocalized with both voltage-gated sodium channels and ankyrin G at the adult rat neuromuscular junction.
J. Cell Biol
140
, 675
–684
Zhang
Z.
, Devarajan
P.
, Dorfman
A. L.
, Morrow
J. S.
(1998
). Structure of the ankyrin-binding domain of alpha-Na,K-ATPase.
J. Biol. Chem
273
, 18681
–18684
Zhou
D.
, Birkenmeier
C. S.
, Williams
M. W.
, Sharp
J. J.
, Barker
J. E.
, Bloch
R. J.
(1997
). Small, membrane-bound, alternatively spliced forms of ankyrin 1 associated with the sarcoplasmic reticulum of mammalian skeletal muscle.
J. Cell Biol
136
, 621
–631
Zhou
D.
, Ursitti
J. A.
, Bloch
R. J.
(1998
). Developmental expression of spectrins in rat skeletal muscle.
Mol. Biol. Cell
9
, 47
–61
This content is only available via PDF.
© 2001 by Company of Biologists
2001
(update)-MotorCFPExtended.jpg?versionId=3964)
-bioRxiv.jpg?versionId=3964)
(update)-CSTW.png?versionId=3964)
-FocalPlaneNetwork.png?versionId=3964)
-Mole.png?versionId=3964)
