Nucleic Acids Research, Vol. 18, No. 18 5547
k.) 1990 Oxford University Press
Nucleotide sequence of the tolC gene of Escherichia coli Hironori Niki, Ryu Imamura, Teru Ogura and Sota Hiraga Department of Molecular Genetics, Institute for Medical Genetics, Kumamoto University Medical School, Kumamoto 862, Japan Submitted August 17, 1990
EMBL accession no. X54049
The mutants of the tolC gene encoding an outer membrane protein exhibit pleiotrophic effects, for example on expression of outer membrane proteins (1) and partition of chromosome (2). We show here our result of sequencing of the BglI-PvuIl DNA segment (2096 bp) including the tolC (mukkA) gene, which was cloned from the wild type W3 110 strain of E. coli. The coding region has 1485 bp, specifying 495 amino acids. A DNA sequence of tolC has already been described (3), however we have found that the sequence by them is inconsistent with our result in the large central part of the gene including the Pvull (position 908) and KpnI (position 1178) sites, which are present in both the cleavage maps of the tolC segments cloned by them and by us and also in the cleavage map of Kohara et al. (4), however absent in the sequence described by Hackett and Reeves
ACKNOWLEDGEMENTS We thank Dr P.Reeves for sending their tolC clones. This work was supported by grants from the Ministry of Education, Science and Culture of Japan. REFERENCES 1. Morona,R. and Reeves,P. (1982) J. Bacteriol. 150, 1016-1023. 2. Hiraga,S., Niki,H., Ogura,T., Ichinose,C., Mori,H., Ezaki,B. and Jaffe,A. (1989) J. Bacteriol. 170, 1496-1505. 3. Hackett,J. and Reeves,P. (1983) Nucd. Acids Res. 11, 6487-6495. 4. Kohara,Y., Akiyama,K. and Isono,K. (1987) Cell 50, 495-508.
(3). AGATCTAATGAAAAAAAGCCGCGATAAAGTGTTTCTCGTGCAATAATTTCTACATCGTTTTTGCCAAATGTAACGGGCAGGTTGTCTGGCTTAAGCATTG TTAATGTCCTGGCACTAATAGTGAATTAAATGTGAATTTCAGCGACGTTTGACTGCCGTTTGAGCAGTCATGTGTTAAATTGAGGCACATTAACGCCCTA TGGCACGTAACGCCAACCTTTTGCGGTCAGGCTTCTGCTAGAATCCGCAATAATTTTACAGTTTGATCGCGCTAAATACTGCTTCACCACAAGGAATGCA Q
M
AATGAAGAAATTGCTCCCCATTCTTATCGGCCTGAGCCTTTCTGGGTTCAGTTCGTTGAGCCAGGCCGAGAACCTGATGCAAGTTTATCAGCAAGCACGC M
K
K
L
L
P
L
I
I
G
L
S
L
S
G
S
F
S
L
S
Q
A
E
N
L
M
Q
V
Y
Q
Q
A
S
P
N
E
L
K
R
A
S
A
R
D
D
A
A
E
F
K
I
N
E
A
R
S
P
L
L
P
Q
L
G
L
D
Y
T
Y
S
N
G
Y
R
D
A
N
G
I
N
S
N
A
T
S
A
S
L
Q
L
T
Q
S
I
F
D
500
G
GTGCAGATTACACCTATAGCAACGGCTACCGCGACGCGAACGGCATCAACTCTAACGCGACCAGTGCGTCCTTGCAGTTAACTCAATCCATTTTTGATAT A
400
R
CTTAGTAACCCGGAATTGCGTAAGTCTGCCGCCGATCGTGATGCTGCCTTTGAAAAAATTAATGAAGCGCGCAGTCCATTACTGCCACAGCTAGGTTTAG L
100 200 300
600
M
GTCGAAATGGCGTGCGTTAACGCTGCAGGAAAAAGCAGCAGGGATTCAGGACGTCACGTATCAGACCGATCAGCAAACCTTGATCCTCAACACCGCGACC 700 S K W R A L T L Q E K A A G I Q D V T Y Q T D Q Q T L I L N T A T GCTTATTTCAACGTGTTGAATGCTATTGACGTTCTTTCCTATACACAGGCACAAAAAGAAGCGATCTACCGTCAATTAGATCAAACCACCCAACGTTTTA 800 A Y F N V L N A I D V L S Y T Q A Q K E A I Y R Q L D Q T T Q R F N ACGTGGGCCTGGTAGCGATCACCGACGTGCAGAACGCCCGCGCACAGTACGATACCGTGCTGGCGAACGAACTGACCGCACGTAATAACCTTGATAACGC 900 V G L V A I T D V Q N A R A Q Y D T V L A N E L T A R N N L D N A GGTAGAGCAGCTGCGCCAGATCACCGGTAACTACTATCCGGAACTGGCTGCGCTGAATGTCGAAAACTTTAAAACCGACAAACCACAGCCGGTTAACGCG 1000 V E Q L R Q I T G N Y Y P E L A A L N V E N F K T D K P Q P V N A CTGCTGAAAGAAGCCGAAAAACGCAACCTGTCGCTGTTACAGGCACGCTTGAGCCAGGACCTGGCGCGCGAGCAAATTCGCCAGGCGCAGGATGGTCACT 1100 L L K E A E K R N L S L L Q A R L S Q D L A R E Q I R Q A Q D G H L TACCGACTCTGGATTTAACGGCTTCTACCGGGATTTCTGACACCTCTTATAGCGGTTCGAAAACCCGTGGTGCCGCTGGTACCCAGTATGACGATAGCAA 1200 P T L D L T A S T G I S D T S Y S G S K T R G A A G T Q Y D D S N TATGGGCCAGAACAAAGTTGGCCTGAGCTTCTCGCTGCCGATTTATCAGGGCGGAATGGTTAACTCGCAGGTGAAACAGGCACAGTACAACTTTGTCGGT 1300 M G Q N K V G L S F S L P I Y Q G G M V N S Q V K Q A Q Y N F V G GCCAGCGAGCAACTGGAAAGTGCCCATCGTAGCGTCGTGCAGACCGTGCGTTCCTCCTTCAACAACATTAATGCATCTATCAGTAGCATTAACGCCTACA 1400 A S E Q L E S A H R S V V Q T V R S S F N N I N A S I S S I N A Y K AACAAGCCGTAGTTTCCGCTCAAAGCTCATTAGACGCGATGGAAGCGGGCTACTCGGTCGGTACGCGTACCATTGTTGATGTGTTGGATGCGACCACCAC 1500 Q A V V S A Q S S L D A M E A G Y S V G T R T I V D V L D A T T T GTTGTACAACGCCAAGCAAGAGCTGGCGAATGCGCGTTATAACTACCTGATTAATCAGCTGAATATTAAGTCAGCTCTGGGTACGTTGAACGAGCAGGAT 1600 L Y N A K Q E L A N A R Y N Y L I N Q L N I K S A L G T L N E Q D CTGCTGGCACTGAACAATGCGCTGAGCAAACCGGTTTCCACTAATCCGGAAAACGTTGCACCGCAAACGCCGGAACAGAATGCTATTGCTGATGGTTATG 1700 L L A L N N A L S K P V S T N P E N V A P Q T P E Q N A I A D G Y A CGCCTGATAGCCCGGCACCAGTCGTTCAGCAAACATCCGCACGCACTACCACCAGTAACGGTCATAACCCTTTCCGTAACTGATGACGACGACGGGGCTT 1800 P D S P A P V V Q Q T S A R T T T S N G H N P F R N * CGGCCCCGTCTGAACGTAAGGCAACGTAAAGATACGGGTTATCTGCCGCATTCTTCCCCCTTCTCGCTTCAATTTCGACCAGCCATCCTCTATTCTGATG 1900
GGTATTTACCACTGGTCCCGGAAGACAAAAATGAAACGGACAAAATCCATACGCCACGCATCGTTCCGCAAAAACTGGAGCGCACGCCATCTGACACCAG 2000 2096 TCGCTCTCGCGGTTGCCACTGTTTTTATGCTGGCTGGCTGTGAAAAGAGTGATGAAACAGTGTCTCTCTATCAAAATGCTGACGACTGTTCAGCTG
k.) 1990 Oxford University Press
Nucleotide sequence of the tolC gene of Escherichia coli Hironori Niki, Ryu Imamura, Teru Ogura and Sota Hiraga Department of Molecular Genetics, Institute for Medical Genetics, Kumamoto University Medical School, Kumamoto 862, Japan Submitted August 17, 1990
EMBL accession no. X54049
The mutants of the tolC gene encoding an outer membrane protein exhibit pleiotrophic effects, for example on expression of outer membrane proteins (1) and partition of chromosome (2). We show here our result of sequencing of the BglI-PvuIl DNA segment (2096 bp) including the tolC (mukkA) gene, which was cloned from the wild type W3 110 strain of E. coli. The coding region has 1485 bp, specifying 495 amino acids. A DNA sequence of tolC has already been described (3), however we have found that the sequence by them is inconsistent with our result in the large central part of the gene including the Pvull (position 908) and KpnI (position 1178) sites, which are present in both the cleavage maps of the tolC segments cloned by them and by us and also in the cleavage map of Kohara et al. (4), however absent in the sequence described by Hackett and Reeves
ACKNOWLEDGEMENTS We thank Dr P.Reeves for sending their tolC clones. This work was supported by grants from the Ministry of Education, Science and Culture of Japan. REFERENCES 1. Morona,R. and Reeves,P. (1982) J. Bacteriol. 150, 1016-1023. 2. Hiraga,S., Niki,H., Ogura,T., Ichinose,C., Mori,H., Ezaki,B. and Jaffe,A. (1989) J. Bacteriol. 170, 1496-1505. 3. Hackett,J. and Reeves,P. (1983) Nucd. Acids Res. 11, 6487-6495. 4. Kohara,Y., Akiyama,K. and Isono,K. (1987) Cell 50, 495-508.
(3). AGATCTAATGAAAAAAAGCCGCGATAAAGTGTTTCTCGTGCAATAATTTCTACATCGTTTTTGCCAAATGTAACGGGCAGGTTGTCTGGCTTAAGCATTG TTAATGTCCTGGCACTAATAGTGAATTAAATGTGAATTTCAGCGACGTTTGACTGCCGTTTGAGCAGTCATGTGTTAAATTGAGGCACATTAACGCCCTA TGGCACGTAACGCCAACCTTTTGCGGTCAGGCTTCTGCTAGAATCCGCAATAATTTTACAGTTTGATCGCGCTAAATACTGCTTCACCACAAGGAATGCA Q
M
AATGAAGAAATTGCTCCCCATTCTTATCGGCCTGAGCCTTTCTGGGTTCAGTTCGTTGAGCCAGGCCGAGAACCTGATGCAAGTTTATCAGCAAGCACGC M
K
K
L
L
P
L
I
I
G
L
S
L
S
G
S
F
S
L
S
Q
A
E
N
L
M
Q
V
Y
Q
Q
A
S
P
N
E
L
K
R
A
S
A
R
D
D
A
A
E
F
K
I
N
E
A
R
S
P
L
L
P
Q
L
G
L
D
Y
T
Y
S
N
G
Y
R
D
A
N
G
I
N
S
N
A
T
S
A
S
L
Q
L
T
Q
S
I
F
D
500
G
GTGCAGATTACACCTATAGCAACGGCTACCGCGACGCGAACGGCATCAACTCTAACGCGACCAGTGCGTCCTTGCAGTTAACTCAATCCATTTTTGATAT A
400
R
CTTAGTAACCCGGAATTGCGTAAGTCTGCCGCCGATCGTGATGCTGCCTTTGAAAAAATTAATGAAGCGCGCAGTCCATTACTGCCACAGCTAGGTTTAG L
100 200 300
600
M
GTCGAAATGGCGTGCGTTAACGCTGCAGGAAAAAGCAGCAGGGATTCAGGACGTCACGTATCAGACCGATCAGCAAACCTTGATCCTCAACACCGCGACC 700 S K W R A L T L Q E K A A G I Q D V T Y Q T D Q Q T L I L N T A T GCTTATTTCAACGTGTTGAATGCTATTGACGTTCTTTCCTATACACAGGCACAAAAAGAAGCGATCTACCGTCAATTAGATCAAACCACCCAACGTTTTA 800 A Y F N V L N A I D V L S Y T Q A Q K E A I Y R Q L D Q T T Q R F N ACGTGGGCCTGGTAGCGATCACCGACGTGCAGAACGCCCGCGCACAGTACGATACCGTGCTGGCGAACGAACTGACCGCACGTAATAACCTTGATAACGC 900 V G L V A I T D V Q N A R A Q Y D T V L A N E L T A R N N L D N A GGTAGAGCAGCTGCGCCAGATCACCGGTAACTACTATCCGGAACTGGCTGCGCTGAATGTCGAAAACTTTAAAACCGACAAACCACAGCCGGTTAACGCG 1000 V E Q L R Q I T G N Y Y P E L A A L N V E N F K T D K P Q P V N A CTGCTGAAAGAAGCCGAAAAACGCAACCTGTCGCTGTTACAGGCACGCTTGAGCCAGGACCTGGCGCGCGAGCAAATTCGCCAGGCGCAGGATGGTCACT 1100 L L K E A E K R N L S L L Q A R L S Q D L A R E Q I R Q A Q D G H L TACCGACTCTGGATTTAACGGCTTCTACCGGGATTTCTGACACCTCTTATAGCGGTTCGAAAACCCGTGGTGCCGCTGGTACCCAGTATGACGATAGCAA 1200 P T L D L T A S T G I S D T S Y S G S K T R G A A G T Q Y D D S N TATGGGCCAGAACAAAGTTGGCCTGAGCTTCTCGCTGCCGATTTATCAGGGCGGAATGGTTAACTCGCAGGTGAAACAGGCACAGTACAACTTTGTCGGT 1300 M G Q N K V G L S F S L P I Y Q G G M V N S Q V K Q A Q Y N F V G GCCAGCGAGCAACTGGAAAGTGCCCATCGTAGCGTCGTGCAGACCGTGCGTTCCTCCTTCAACAACATTAATGCATCTATCAGTAGCATTAACGCCTACA 1400 A S E Q L E S A H R S V V Q T V R S S F N N I N A S I S S I N A Y K AACAAGCCGTAGTTTCCGCTCAAAGCTCATTAGACGCGATGGAAGCGGGCTACTCGGTCGGTACGCGTACCATTGTTGATGTGTTGGATGCGACCACCAC 1500 Q A V V S A Q S S L D A M E A G Y S V G T R T I V D V L D A T T T GTTGTACAACGCCAAGCAAGAGCTGGCGAATGCGCGTTATAACTACCTGATTAATCAGCTGAATATTAAGTCAGCTCTGGGTACGTTGAACGAGCAGGAT 1600 L Y N A K Q E L A N A R Y N Y L I N Q L N I K S A L G T L N E Q D CTGCTGGCACTGAACAATGCGCTGAGCAAACCGGTTTCCACTAATCCGGAAAACGTTGCACCGCAAACGCCGGAACAGAATGCTATTGCTGATGGTTATG 1700 L L A L N N A L S K P V S T N P E N V A P Q T P E Q N A I A D G Y A CGCCTGATAGCCCGGCACCAGTCGTTCAGCAAACATCCGCACGCACTACCACCAGTAACGGTCATAACCCTTTCCGTAACTGATGACGACGACGGGGCTT 1800 P D S P A P V V Q Q T S A R T T T S N G H N P F R N * CGGCCCCGTCTGAACGTAAGGCAACGTAAAGATACGGGTTATCTGCCGCATTCTTCCCCCTTCTCGCTTCAATTTCGACCAGCCATCCTCTATTCTGATG 1900
GGTATTTACCACTGGTCCCGGAAGACAAAAATGAAACGGACAAAATCCATACGCCACGCATCGTTCCGCAAAAACTGGAGCGCACGCCATCTGACACCAG 2000 2096 TCGCTCTCGCGGTTGCCACTGTTTTTATGCTGGCTGGCTGTGAAAAGAGTGATGAAACAGTGTCTCTCTATCAAAATGCTGACGACTGTTCAGCTG
