Gene, 112 (1992) 265-266 © 1992 Elsevier Science Publishers B.V. All fights reserved. 0378-1119/92/$05.00
265
GENE 06346
Brief Note Nucleotide sequences of the rabbit ~-smooth-muscle and p-non-muscle actin mRNAs* (Contractile; isoforms; recombinant DNA; uterus)
David E. Harris a, David M. Warshawa and Muthu Periasamy a,b a Departments of Physiology and Biophysics, and b ~,ficrobiology and Molecular Genetics, University of Vermont, Burlington, VT 05405 (U.S.A.) Received by R. Padmanabhan: 21 October 1991 Accepted: 27 November 1991 Received at publishers: 7 January 1992
SUMMARY
We report the characterization of cDNAs encoding two actin isoforms (0c-smooth muscle and p-non-muscle) from the rabbit uterus. These isoforms have extensive amino acid and nucleotide sequence homology to the corresponding mouse and human isoforms.
Actin is a highly conserved contractile and cytoskeletal protein ubiquito~:s in ¢ukaryotes (Vandekerckhove and Weber, 1984). Six actin isoforms have been identified in mammals (Vandekerck~,ove and Weber, 1978) however their functional £~ifferences remain obscure. Towards understanding the structure-function relationship of actin isoforms, cDNAs encoding two actin isoforms from the rabbit were cloned and sequenced. A cDNA library constructed from the poly(A)+mRNA of rabbit uterus (Lytton et al., 1989) was screened using a 500-bp segment of the rat skeletal muscle actin coding sequence as a probe. Two unique full length clones (RSMA2,4 and RSMA-7A) were identified. Both clones were sequenced by constructing nested deletions with exonuclease III (Promega, Madison, WI) and sequencing by the
dideoxy chain termination method (Sanger et al., 1977) (Fig. la and b). The deduced aa sequence of clone RSMA-2A identified it as 0c-smooth muscle actin, while that of RSMA-7A showed it to be/1-non-muscle actin (Vandekerckhove and Weber, 1979). These two rabbit actin isoforms show extensive nt homology to mouse and human (Table I). The deduced aa sequence of the rabbit 0c-smooth muscle actin was identical to that of mouse and differed from the human form only at aa TM (rabbit = S TM, human = W234). The deduced aa sequence of rabbit//-non-muscle actin differed from both the mouse and human at aa ~°~ (mouse and human = H l°l, rabbit = Rl°m). TABLE I Actin nt homology Rabbit
Correspondence to: Dr. D.E. Harris, Department of Physiology and Biophysics, University of Vermont, Burlington, VT 05405 (U.S.A.) Tel. (802)656-3411; Fax (802)656-0747 * On request, the authors will supply detailed experimental evidence for the conclusions reached in this Brief Note. Abbreviations: aa, amino acid(s); bp, base pair(s); h, human; kb, kilobase(s) or 1000 bp; m, murine (mouse); nt, nucleotide(s); r, rabbit.
a-smooth muscle //-non muscle
Percent homology to: a Mouse (m)
Human (h)
88 82
93 85
a The nt sequences coding for m and h isoforms were obtained from GenBank (accession numbers X13839 and M10277 for h, and X13297 and X03672 for m.
266
-57 1 3 2
63 22 123 42 183 62 243
82 3N3
102 363
122 423 142 483 162 543 182 603 222 663 242 723 262 783 282 843 282 903 302 963 322 1023
342 1083 362 1143 1203 1263
GGCACGAGGGAGAAGCCCAGCCCAGCACTGTCAGGAATCCCGTGAAGCAGCTGCAGCTAT M _GTGTGAGGAAGAG GACAGCACCG C C C T T G T G T G T G A C A A T G G CTC C G G G C T C T G T A A G G C C
E
E
E
D
S
T
A
L
V
C
D
N
G
S
G
L
C
K
A
TGGCTTTGCCGGAGACGATG CTCCAAGAG C T G T T T T C C C A T C C A T T G T G G G A C G T C C C A G G F A G D D A P R A V F P S I V G R P R GCACCAGGGGGTGATGGTGGGAATGGGGCAAAAAGACAGCTACGTGGGTGATGAAGCGCA H Q G V M V G M G Q K D S Y V G D E A Q GAGCAAAAGAGGAATCCTGACCTTGAAGTACCCGATCGAACATGGCATCATCACCAACTG S K R G I L T L K ¥ P I E H G I I T N W GGACGACATGGAAAAGATCTGGCACCACTCCTTCTACAATGAGCTTCGCGTTGCCCCTGA D D M E K I W H H S F ¥ N E L R V A P E AGAACATCCAACCCTGTTGACTGAGGCACCGCTGAACCCCAAAGCCAACCGGGAGAAAAT E H P T L L T E A P L N P K A N R E K M G A C C C A G A T T A T G T T T G A G A C T T T C A A T G T C C C A G C C A T G T A C G T G G C T A T T C A G G CGGT T Q I M F E T F N V P A H ¥ V A I Q A V GCTGTCCCTCTATGCCTCTGGACGTACAACTGGCATTGTGCTGGACTCTGGAGATGGTGT L S L ¥ A S G R T T G I V L D S G D G V CACCCACAACGTGCCCATCTATGAGGG C T A T G C C C T G C C C C A T G C C A T C A T G C G T C T G G A T H N V P I ¥ E G Y A L P H A I M R L D CCTGG CAGG C C G A G A T C T C A C T G A C T A C C T C A T G A A G A T C C T G A C T G A G C G T G G C T A T T C L A G R D L T D Y L M K I L T E R G Y S CTTCGTGACTACTG C T G A A C G T G A G A T T G T C C G G G A C A T C A A A G A G A A A T T G T G CTATGT F V T T A E R E I V R P I K E K L C Y V CGCTCTGGACTTTGAAAACGAGATGGCCACTGCTGCATCCTCCTCCTCCCTCGAGAAGAG A L D P E N E M A T A A S S S S L E K S CTATGAGCTGCCTGACGGGeAGGTGATCACCATCGGGAACGAACGCTTCCGCTGCCCAGA Y E L P D G Q V I T I G N E R F R C P E GACCCTGTTCCAGCCCTC C T T C A T C G G G A T G G A A T C T G C T G G C A T C C A T G A A A C C A C C T A T L F Q P S F I G M E S A G I H E T T Y CAACAGCATCATGAAGTGTGACATCGACATCAGGAAGGACCTCTATGCTAACAATGTGCT N S I M K C D I D I R K D L ¥ A N N V L CTCAGGGGGCACCACCATGTACCCTGGCATTGCTGACCGTATGCAGAAGGAAATCACGGC S G G T T M Y P G I A D R M Q K E I T A CCTAGCTCCCAGCACCATGAAGATCAAGATAATTGCCCCTCCAGAG CGCAAATACTCCGT L A P S T M K I K I I A P P E R K ¥ S V CTGGATCGGCGGCTCCATCCTGGCCTCTCTGTCCACCTTTCAGCAGATGTGGATCAGCAA W I G G S I L A S L S T F Q Q M W I S K ACAGGAGTACGATGAAGCCGGGCCCTCCATTGTCCACCGCAAATGCTTCTAAGTCACTTC Q E ¥ D E A G P S I V H R K C F 377 CCTGCTCTGTTTCTAGCCCACAACTGTGAATGTGTTGTGGAATAATGCCTTCAATTCCTT TCCAAGTCATGCCTATCCAAAGCTCTGACTCATTACCTATGTATTTTTTAATAA~TCTGA AATATGCTACCAGTCAA 1279
-74 GGCACGAGCCAGCTGCCGACCACCGCACCTC3CCCAGCCGCTCCGCCGCCGGCCCACACT -14 CCCGCTCAGCTCACCATGGATGACGATATCGCCGCGCTCGTGGTCGAC~TGGCTCCGGC 1 M D D D I A A L V V D N G S G 46 A T G T G C ~ G G C C G G C T T C G C G G G C G A C G A T G C C C C C C G G G C C G T ~ T C C C C T C C A T C G T G 1 6 M C K A G F A G D D A P R A V F P S I V 106 G G G C G T C C G C G G C A C C A G G G C G T G A T G G T G G G C A T G G G C C A G ~ G G A C T C G T A C G T G G G C 3 6 G R P R H Q G V N V G H G Q K D S Y V G 166 G A C G A G G C C C A G A G C ~ G C G T G G C A T C C T G A C G C T C ~ G T A C C C G A T C G A G C A C G G C A T C 5 6 D E A Q S K R G I L T L K Y P I E H G I 226 G T G A C C ~ C T G G G A C G A C A T G G A G ~ G A T C T G G C A C C A C A C C T T C T A C ~ C G A G C T T C G C 7 6 V T N W D D M E K I W H H T F Y N E L R 286 G T G G C G C C C G A G G ~ C G C C C G G T G C T G C T G A C C G A G G C C C C C C T G ~ C C C C ~ G G C C ~ C 9 6 V A P E E R P V L L T E A P L N P K A N 346 C G C G A G ~ G A T G A C C C A G A T C A T G T T C G A G A C G T T C ~ C A C G C C G G C C A T G T A C G T G G C C 1 1 6 R E K M T Q I M F E T F N T P A M Y V A 406 A T C C A G G C C G T G C T G T C C C T G T A C G C C T C T G G C C G C A C C A C G G G C A T C G T G A T G G A C T C T 1 3 6 I Q A V L S L Y A S G R T T G I V , ~ M D S 466 G G C G A C G G G G T C A C C C A C A C G G T G C C C A T C T A C G A G C G C T A C G C G C T G C C G C A C G C C A T C 1 5 6 G D G V T H T V P I Y E G Y A L P H A I , 526 CTGCGTCTGGACCTGGCCGGCCGGGACCTGACCGACTACCTCATG~GATCCTCACGGAG 1 7 6 L R L D L A G R D L T D Y L H K I L T E 586 C G C G G C T A C A G C T T C A C C A C C A C G G C C G A G C G C G A G A T C G T G C G G G A C A T C ~ G G A G ~ G 1 9 6 R G Y S F T T T A E R E I V R D I K E K 646 C T G T G C T A C G T G G C G C T G G A C T T C G A G C A G G A G A T G G C C A C G G C C G C G T C C T C C T C C T C C 2 1 6 L C Y V A L D F E Q E M A T A A S S S S 706 C T G G A G ~ G A G C T A C G A G C T G C C C G A C G G C C A G G T C A T C A C C A T C G G C ~ C G A G C G C T T C 2 3 6 L E K S Y E L P D G Q V I T I G N E R F 766 CGTTGCCCCGAGGCGCTCTTCCAGCCCTCCTTCCTGGGCATGGAGTCGTGTGGCATCCAC 2 5 6 R C P E A L F Q P S F L G M E S C G I H 826 G A G A C C A C C T T C ~ C T C G A T C A T G ~ G T G C G A C G T G G A C A T C C G C ~ G G A C C T G T A C G C C 2 7 6 E T T F N S I M K C D V D I R K D L Y A 886 ~ C A C G G T G C T G T C G G G C G G C A C C A C C A T G T A C C C C G G C A T C G C C G A C C G C A T G C A G A A G , 2 9 6 N T V L S G G T T M Y P G I A D R M Q K 946 G A G A T C A C C G C C C T G G C G C C C A G C A C C A T G ~ G A T C ~ G A T C A T C G C G C C C C C G G A G C G C 3 1 6 E I T A L A P S T M K I K I I A P P E R 1006 ~ G T A C T C G G T G T G G A T C G G C G G C T C C A T C C T G G C C T C G C T C T C C A C C T T C C A G C A G A T G 3 3 6 K Y S V W I G G S I L A S L S T F Q Q M 1066 T G G A T C A G C ~ G C A G G A G T A T G A C G A G T C G G G C C C C T C C A T C G T G C A C C G C ~ G T G C T T C 3 5 6 W I S K Q E Y D E S G P S I V H R K C F 3 7 5 1126 T A G G C G G A C T G T T A G A G C C A G T G G C G G G A C A C C C T C T C T C G A C G A A A C C T ~ C G G C G C A G 1186 A A A C G A G A C G A G A T T G G C A T G G C T T T A T T C G T G T T T T T T T T T T T T T T C T T T T T T T G T T T T 1246 T T T T T T T T T G T T T T G G C G C ~ T T G A C T C A G G A T T T A A A A A C T G G ~ C G G T G ~ G G T G A C A 1306 G C A G T C G T T G G A G C G A G C A T C C C C A A A G T C C C C A A A G T T C T G C ~ T G T G G C C G A G G A C T T 1366 T G A T T G T A C A T T G T T T T T T T T T T T T T C T T T T G A T A G T C A T T C C A A A T A T C G T G A G A T G C C 1426 A T G T G A C G G ~ G C C C C C G C C C C A C T C C T G C C A A G G A G ~ G C G C A G T C C T C G C C G C C C G G G 1486 G C C A C C C A G G G G A G G C G G C A G C A T C G C A T T T G T G T A A A T T A T G T ~ T G C A A A T T T T T T ~ 1546 A T C T T C G C C T T ~ T A C T T T T T T T ~ T T T T C T T T T T A T T T T G ~ T G G T C A G C C A T C G T G G C 1606 CCCCCCTTTTTTGTCCCCC~CTTGAGATGTATG~GGCTTTTGGTCTCCCTGGGAGTGG 1666 G T T G A G G T G T G G A G G C A G C C A G G G C T T C C C T G T A C A C T G A C G A G A C C A G T T C A A T A A A A G 1726 T G C A C A C C T G ~ C A C A A ~ 1764
Fig. 1. The nt sequence encoding rabbit ~-smooth muscle actin (RSMA-2A) (A), and rabbit/l-non-muscle actin (RSMA-TA) (B). The start, stop and polyadenylation signals are underlined. EMBL Data Libr~y accession Nos. are X60732 and X60733, respectively. The deduced aa, placed below the nt sequence, are aligned with the first nt of each codon.
ACKNOWLEDGEMENTS This work was supported by grants from the National Institutes of Health (AR34872 to D . M . W . ; R29-38355 to M.P.). D . M . W . and M.P. are Established Investigators o f the American Heart Association.
REFERENCES Lytton, J., Zarain-Herzber, A., Periasamy, M, and MacLennan, D.H,: Molecular cloning of the mammalian smooth muscle sarco(endo)plasmic reticulum Ca2+-ATPase. J, Biol, Chem. 264 (1989) 70597065
Sanger, F., Nicklen, S. and Coulson A.R.: DNA sequencing with chainterminating inhibitors. Proc. Natl. Acad. Sci. USA 74 (1977) 54635467. Vandekerckhove, J. and Weber, K: At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal peptide. J. Mol. Biol. 126 (1978) 783-802. Vandekerckhove, J. and Weber, K: The complete amino acid sequences of actins from bovine aorta, bovine heart, bovine fast skeletal muscle, and rabbit slow skeletal muscle. Differentiation 14 (1979) 123-133, Vandekerckhove, J. and Weber, K.: Chordate muscle actins differ distinctly from invertebrate muscle actins. J. Mol. Biol. 179 (1984) 391413.
265
GENE 06346
Brief Note Nucleotide sequences of the rabbit ~-smooth-muscle and p-non-muscle actin mRNAs* (Contractile; isoforms; recombinant DNA; uterus)
David E. Harris a, David M. Warshawa and Muthu Periasamy a,b a Departments of Physiology and Biophysics, and b ~,ficrobiology and Molecular Genetics, University of Vermont, Burlington, VT 05405 (U.S.A.) Received by R. Padmanabhan: 21 October 1991 Accepted: 27 November 1991 Received at publishers: 7 January 1992
SUMMARY
We report the characterization of cDNAs encoding two actin isoforms (0c-smooth muscle and p-non-muscle) from the rabbit uterus. These isoforms have extensive amino acid and nucleotide sequence homology to the corresponding mouse and human isoforms.
Actin is a highly conserved contractile and cytoskeletal protein ubiquito~:s in ¢ukaryotes (Vandekerckhove and Weber, 1984). Six actin isoforms have been identified in mammals (Vandekerck~,ove and Weber, 1978) however their functional £~ifferences remain obscure. Towards understanding the structure-function relationship of actin isoforms, cDNAs encoding two actin isoforms from the rabbit were cloned and sequenced. A cDNA library constructed from the poly(A)+mRNA of rabbit uterus (Lytton et al., 1989) was screened using a 500-bp segment of the rat skeletal muscle actin coding sequence as a probe. Two unique full length clones (RSMA2,4 and RSMA-7A) were identified. Both clones were sequenced by constructing nested deletions with exonuclease III (Promega, Madison, WI) and sequencing by the
dideoxy chain termination method (Sanger et al., 1977) (Fig. la and b). The deduced aa sequence of clone RSMA-2A identified it as 0c-smooth muscle actin, while that of RSMA-7A showed it to be/1-non-muscle actin (Vandekerckhove and Weber, 1979). These two rabbit actin isoforms show extensive nt homology to mouse and human (Table I). The deduced aa sequence of the rabbit 0c-smooth muscle actin was identical to that of mouse and differed from the human form only at aa TM (rabbit = S TM, human = W234). The deduced aa sequence of rabbit//-non-muscle actin differed from both the mouse and human at aa ~°~ (mouse and human = H l°l, rabbit = Rl°m). TABLE I Actin nt homology Rabbit
Correspondence to: Dr. D.E. Harris, Department of Physiology and Biophysics, University of Vermont, Burlington, VT 05405 (U.S.A.) Tel. (802)656-3411; Fax (802)656-0747 * On request, the authors will supply detailed experimental evidence for the conclusions reached in this Brief Note. Abbreviations: aa, amino acid(s); bp, base pair(s); h, human; kb, kilobase(s) or 1000 bp; m, murine (mouse); nt, nucleotide(s); r, rabbit.
a-smooth muscle //-non muscle
Percent homology to: a Mouse (m)
Human (h)
88 82
93 85
a The nt sequences coding for m and h isoforms were obtained from GenBank (accession numbers X13839 and M10277 for h, and X13297 and X03672 for m.
266
-57 1 3 2
63 22 123 42 183 62 243
82 3N3
102 363
122 423 142 483 162 543 182 603 222 663 242 723 262 783 282 843 282 903 302 963 322 1023
342 1083 362 1143 1203 1263
GGCACGAGGGAGAAGCCCAGCCCAGCACTGTCAGGAATCCCGTGAAGCAGCTGCAGCTAT M _GTGTGAGGAAGAG GACAGCACCG C C C T T G T G T G T G A C A A T G G CTC C G G G C T C T G T A A G G C C
E
E
E
D
S
T
A
L
V
C
D
N
G
S
G
L
C
K
A
TGGCTTTGCCGGAGACGATG CTCCAAGAG C T G T T T T C C C A T C C A T T G T G G G A C G T C C C A G G F A G D D A P R A V F P S I V G R P R GCACCAGGGGGTGATGGTGGGAATGGGGCAAAAAGACAGCTACGTGGGTGATGAAGCGCA H Q G V M V G M G Q K D S Y V G D E A Q GAGCAAAAGAGGAATCCTGACCTTGAAGTACCCGATCGAACATGGCATCATCACCAACTG S K R G I L T L K ¥ P I E H G I I T N W GGACGACATGGAAAAGATCTGGCACCACTCCTTCTACAATGAGCTTCGCGTTGCCCCTGA D D M E K I W H H S F ¥ N E L R V A P E AGAACATCCAACCCTGTTGACTGAGGCACCGCTGAACCCCAAAGCCAACCGGGAGAAAAT E H P T L L T E A P L N P K A N R E K M G A C C C A G A T T A T G T T T G A G A C T T T C A A T G T C C C A G C C A T G T A C G T G G C T A T T C A G G CGGT T Q I M F E T F N V P A H ¥ V A I Q A V GCTGTCCCTCTATGCCTCTGGACGTACAACTGGCATTGTGCTGGACTCTGGAGATGGTGT L S L ¥ A S G R T T G I V L D S G D G V CACCCACAACGTGCCCATCTATGAGGG C T A T G C C C T G C C C C A T G C C A T C A T G C G T C T G G A T H N V P I ¥ E G Y A L P H A I M R L D CCTGG CAGG C C G A G A T C T C A C T G A C T A C C T C A T G A A G A T C C T G A C T G A G C G T G G C T A T T C L A G R D L T D Y L M K I L T E R G Y S CTTCGTGACTACTG C T G A A C G T G A G A T T G T C C G G G A C A T C A A A G A G A A A T T G T G CTATGT F V T T A E R E I V R P I K E K L C Y V CGCTCTGGACTTTGAAAACGAGATGGCCACTGCTGCATCCTCCTCCTCCCTCGAGAAGAG A L D P E N E M A T A A S S S S L E K S CTATGAGCTGCCTGACGGGeAGGTGATCACCATCGGGAACGAACGCTTCCGCTGCCCAGA Y E L P D G Q V I T I G N E R F R C P E GACCCTGTTCCAGCCCTC C T T C A T C G G G A T G G A A T C T G C T G G C A T C C A T G A A A C C A C C T A T L F Q P S F I G M E S A G I H E T T Y CAACAGCATCATGAAGTGTGACATCGACATCAGGAAGGACCTCTATGCTAACAATGTGCT N S I M K C D I D I R K D L ¥ A N N V L CTCAGGGGGCACCACCATGTACCCTGGCATTGCTGACCGTATGCAGAAGGAAATCACGGC S G G T T M Y P G I A D R M Q K E I T A CCTAGCTCCCAGCACCATGAAGATCAAGATAATTGCCCCTCCAGAG CGCAAATACTCCGT L A P S T M K I K I I A P P E R K ¥ S V CTGGATCGGCGGCTCCATCCTGGCCTCTCTGTCCACCTTTCAGCAGATGTGGATCAGCAA W I G G S I L A S L S T F Q Q M W I S K ACAGGAGTACGATGAAGCCGGGCCCTCCATTGTCCACCGCAAATGCTTCTAAGTCACTTC Q E ¥ D E A G P S I V H R K C F 377 CCTGCTCTGTTTCTAGCCCACAACTGTGAATGTGTTGTGGAATAATGCCTTCAATTCCTT TCCAAGTCATGCCTATCCAAAGCTCTGACTCATTACCTATGTATTTTTTAATAA~TCTGA AATATGCTACCAGTCAA 1279
-74 GGCACGAGCCAGCTGCCGACCACCGCACCTC3CCCAGCCGCTCCGCCGCCGGCCCACACT -14 CCCGCTCAGCTCACCATGGATGACGATATCGCCGCGCTCGTGGTCGAC~TGGCTCCGGC 1 M D D D I A A L V V D N G S G 46 A T G T G C ~ G G C C G G C T T C G C G G G C G A C G A T G C C C C C C G G G C C G T ~ T C C C C T C C A T C G T G 1 6 M C K A G F A G D D A P R A V F P S I V 106 G G G C G T C C G C G G C A C C A G G G C G T G A T G G T G G G C A T G G G C C A G ~ G G A C T C G T A C G T G G G C 3 6 G R P R H Q G V N V G H G Q K D S Y V G 166 G A C G A G G C C C A G A G C ~ G C G T G G C A T C C T G A C G C T C ~ G T A C C C G A T C G A G C A C G G C A T C 5 6 D E A Q S K R G I L T L K Y P I E H G I 226 G T G A C C ~ C T G G G A C G A C A T G G A G ~ G A T C T G G C A C C A C A C C T T C T A C ~ C G A G C T T C G C 7 6 V T N W D D M E K I W H H T F Y N E L R 286 G T G G C G C C C G A G G ~ C G C C C G G T G C T G C T G A C C G A G G C C C C C C T G ~ C C C C ~ G G C C ~ C 9 6 V A P E E R P V L L T E A P L N P K A N 346 C G C G A G ~ G A T G A C C C A G A T C A T G T T C G A G A C G T T C ~ C A C G C C G G C C A T G T A C G T G G C C 1 1 6 R E K M T Q I M F E T F N T P A M Y V A 406 A T C C A G G C C G T G C T G T C C C T G T A C G C C T C T G G C C G C A C C A C G G G C A T C G T G A T G G A C T C T 1 3 6 I Q A V L S L Y A S G R T T G I V , ~ M D S 466 G G C G A C G G G G T C A C C C A C A C G G T G C C C A T C T A C G A G C G C T A C G C G C T G C C G C A C G C C A T C 1 5 6 G D G V T H T V P I Y E G Y A L P H A I , 526 CTGCGTCTGGACCTGGCCGGCCGGGACCTGACCGACTACCTCATG~GATCCTCACGGAG 1 7 6 L R L D L A G R D L T D Y L H K I L T E 586 C G C G G C T A C A G C T T C A C C A C C A C G G C C G A G C G C G A G A T C G T G C G G G A C A T C ~ G G A G ~ G 1 9 6 R G Y S F T T T A E R E I V R D I K E K 646 C T G T G C T A C G T G G C G C T G G A C T T C G A G C A G G A G A T G G C C A C G G C C G C G T C C T C C T C C T C C 2 1 6 L C Y V A L D F E Q E M A T A A S S S S 706 C T G G A G ~ G A G C T A C G A G C T G C C C G A C G G C C A G G T C A T C A C C A T C G G C ~ C G A G C G C T T C 2 3 6 L E K S Y E L P D G Q V I T I G N E R F 766 CGTTGCCCCGAGGCGCTCTTCCAGCCCTCCTTCCTGGGCATGGAGTCGTGTGGCATCCAC 2 5 6 R C P E A L F Q P S F L G M E S C G I H 826 G A G A C C A C C T T C ~ C T C G A T C A T G ~ G T G C G A C G T G G A C A T C C G C ~ G G A C C T G T A C G C C 2 7 6 E T T F N S I M K C D V D I R K D L Y A 886 ~ C A C G G T G C T G T C G G G C G G C A C C A C C A T G T A C C C C G G C A T C G C C G A C C G C A T G C A G A A G , 2 9 6 N T V L S G G T T M Y P G I A D R M Q K 946 G A G A T C A C C G C C C T G G C G C C C A G C A C C A T G ~ G A T C ~ G A T C A T C G C G C C C C C G G A G C G C 3 1 6 E I T A L A P S T M K I K I I A P P E R 1006 ~ G T A C T C G G T G T G G A T C G G C G G C T C C A T C C T G G C C T C G C T C T C C A C C T T C C A G C A G A T G 3 3 6 K Y S V W I G G S I L A S L S T F Q Q M 1066 T G G A T C A G C ~ G C A G G A G T A T G A C G A G T C G G G C C C C T C C A T C G T G C A C C G C ~ G T G C T T C 3 5 6 W I S K Q E Y D E S G P S I V H R K C F 3 7 5 1126 T A G G C G G A C T G T T A G A G C C A G T G G C G G G A C A C C C T C T C T C G A C G A A A C C T ~ C G G C G C A G 1186 A A A C G A G A C G A G A T T G G C A T G G C T T T A T T C G T G T T T T T T T T T T T T T T C T T T T T T T G T T T T 1246 T T T T T T T T T G T T T T G G C G C ~ T T G A C T C A G G A T T T A A A A A C T G G ~ C G G T G ~ G G T G A C A 1306 G C A G T C G T T G G A G C G A G C A T C C C C A A A G T C C C C A A A G T T C T G C ~ T G T G G C C G A G G A C T T 1366 T G A T T G T A C A T T G T T T T T T T T T T T T T C T T T T G A T A G T C A T T C C A A A T A T C G T G A G A T G C C 1426 A T G T G A C G G ~ G C C C C C G C C C C A C T C C T G C C A A G G A G ~ G C G C A G T C C T C G C C G C C C G G G 1486 G C C A C C C A G G G G A G G C G G C A G C A T C G C A T T T G T G T A A A T T A T G T ~ T G C A A A T T T T T T ~ 1546 A T C T T C G C C T T ~ T A C T T T T T T T ~ T T T T C T T T T T A T T T T G ~ T G G T C A G C C A T C G T G G C 1606 CCCCCCTTTTTTGTCCCCC~CTTGAGATGTATG~GGCTTTTGGTCTCCCTGGGAGTGG 1666 G T T G A G G T G T G G A G G C A G C C A G G G C T T C C C T G T A C A C T G A C G A G A C C A G T T C A A T A A A A G 1726 T G C A C A C C T G ~ C A C A A ~ 1764
Fig. 1. The nt sequence encoding rabbit ~-smooth muscle actin (RSMA-2A) (A), and rabbit/l-non-muscle actin (RSMA-TA) (B). The start, stop and polyadenylation signals are underlined. EMBL Data Libr~y accession Nos. are X60732 and X60733, respectively. The deduced aa, placed below the nt sequence, are aligned with the first nt of each codon.
ACKNOWLEDGEMENTS This work was supported by grants from the National Institutes of Health (AR34872 to D . M . W . ; R29-38355 to M.P.). D . M . W . and M.P. are Established Investigators o f the American Heart Association.
REFERENCES Lytton, J., Zarain-Herzber, A., Periasamy, M, and MacLennan, D.H,: Molecular cloning of the mammalian smooth muscle sarco(endo)plasmic reticulum Ca2+-ATPase. J, Biol, Chem. 264 (1989) 70597065
Sanger, F., Nicklen, S. and Coulson A.R.: DNA sequencing with chainterminating inhibitors. Proc. Natl. Acad. Sci. USA 74 (1977) 54635467. Vandekerckhove, J. and Weber, K: At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal peptide. J. Mol. Biol. 126 (1978) 783-802. Vandekerckhove, J. and Weber, K: The complete amino acid sequences of actins from bovine aorta, bovine heart, bovine fast skeletal muscle, and rabbit slow skeletal muscle. Differentiation 14 (1979) 123-133, Vandekerckhove, J. and Weber, K.: Chordate muscle actins differ distinctly from invertebrate muscle actins. J. Mol. Biol. 179 (1984) 391413.
