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Nucleic Acids Research, Vol. 18, No. 5
DNA sequence of exoenzyme 03, an ADPribosyltransferase encoded by Clostridium botulinum and D phages
C
Michel Popoff, Patrice Boquet1, D.Michael Gill2* and Melvin W.Eklund3 Unite des Anaerobies and 1Unite des Antigenes Bacteriens, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris, France, 2Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111 and 3Northwest and Alaska Fisheries Center, 2725 Montlake Boulevard East, Seattle, WA 98112, USA Submitted January 29, 1990
EMBL accession no. X51464
Exoenzyme C3 ADP-ribosylates the rho group of small GTPbinding proteins (1, 2). Extracellular C3 from C. botulinum type D is a basic protein of approximate size 26,000 and N terminal AYSNTYQEFTNIDQAKAXGNAQYKKY. Oligonucleotides representing this sequence hybridize to C and D botulinal phage DNA. The nucleotide sequence below was derived from cloned fragments of D phage and represents the extracellular protein of 211 amino acid residues Mr 23,546) plus seven N-terminal amino acid residues which imply a precursor protein cleaved at the site shown with an asterisk. The adjacent 5' DNA sequence is unstable in E. coli and has not been cloned. A candidate transcription terminator (AG = -16 kcalorie) is underlined. C phage have the identical coding sequence and two differences
in the 3' untranslated region: A662, T736. The C3 gene is distinct from the C1 or D neurotoxin genes which are also encoded by these phages (3, 4).
REFERENCES 1. Chardin,P., Boquet,P., Madaule,P., Popoff,M.R., Rubin,E.J. and Gill,D.M. (1989) EMBO J. 8, 1087 - 1092. 2. Rubin,E.J., Gil,D.M., Boquet,P. and Popoff,M.R. (1988) Mol. Cell. Biol. 8, 418-426. 3. Eklund,M.W., Poysky,F.T., Reed,S.M. and Smith,C.A. (1971) Science 172, 480-481. 4. Eklund,M.W., Poysky,F.T. and Reed,S.M. (19972) Nature New Biology 235, 16-17.
*
GCT TAT TCC Ala Tyr Ser CM GCA AAA Gln Ala Lys GM GCT ATA Glu Ala Ile MG GGA GTT
Lys Gly Val TCT TTT AAT Ser Phe Asn TAT Tyr GCT Ala AGT Ser
TTA Leu TTT Phe ACT
GGA
Gly GM Glu TCA
Thr Ser
GTA GCA AAA
Val Ala Lys GAA ATG TTG Glu Met Leu GGT AAA CM Gly Lys Gln AGC TAT TAG AAG TAG CTC
*
ATT AAT CAA AAG GCT TAT TCA MT ACT TAC CAG GAG TTT ACT MT ATT CAT Ile Asn Gln Lys Ala Tyr Ser Asn Thr Tyr Gln Glu Phe Thr Asn Ile Asp GCT 'TGG GGT MT GCT CAG TAT AAA MG TAT GGA CTA AGC AAA TCA GAA AAA Ala 'Trp Gly Asn Ala Gln Tyr Lys Lys Tyr Gly Leu Ser Lys Ser Glu Lys GTA 'TCA TAT ACT AM AGC GCT AGT GM ATA AAT GGA AAG CTA AGA CAA MT Val 'Ser Tyr Thr Lys Ser Ala Ser Glu Ile Asn Gly Lys Leu Arg Gln Asn ATC IMAT GGA TTT CCT TCA AAT TTA ATA AAA CAA GTT GAA CTT TTA GAT AAA Ile IAsn Gly Phe Pro Ser Asn Leu Ile Lys Gln Val Glu Leu Leu Asp Lys AAA iATG AAG ACC CCT GM MT ATT ATG TTA TTT AGA GGC GAC GAC CCT GCT Lys IMet Lys Thr Pro Glu Asn Ile Met Leu Phe Arg Gly Asp Asp Pro Ala ACA (GM TTT CM MC ACT CTT CTT MT TCA AAT GGT ACA ATT AAT AM ACG Thr (Glu Phe Gln Asn Thr Leu Leu Asn Ser Asn Gly Thr Ile Asn Lys Thr MG (GCT AAA GCT MG TTT TTA MT AAA GAT AGA CTT GM TAT GGA TAT ATT Lys AAla Lys Ala Lys Phe Leu Asn Lys Asp Arg Leu Glu Tyr Gly Tyr Ile TTA jATG MT GTT TCT CM TTT GCA GGA AGA CCA ATT ATT ACA AAA TTT AAA Leu IMet Asn Val Ser Gln Phe Ala Gly Arg Pro Ile Ile Thr Lys Phe Lys GGC TCA AAG GCA GGA TAT ATT GAC CCT ATT AGT GCT TTT GCA GGA CM CTT Gly Ser Lys Ala Gly Tyr Ile Asp Pro Ile Ser Ala Phe Ala Gly Gln Leu CTT CCT AGA CAT AGT ACT TAT CAT ATA GAC GAT ATG AGA TTG TCT TCT GAT Leu Pro Arg His Ser Thr Tyr His Ile Asp Asp Met Arg Leu Ser Ser Asp ATA ATA ATT ACA GCA ACA ATG ATG GGC ACA GCT ATC MT CCT AAA TM TAG Ile Ile Ile Thr Ala Thr Met Met Gly Thr Ala Ile Asn Pro Lys *** *** CAT GTT MG GM TTG TAT ATA ATT AM TGT MA MG AGT TAC TTT ATA ACA TTT TTA CTA TM GCA AAA ATA AGA CTG CTA TTT ATA CCC MT ATA TGG CM
To whom correspondence should be addressed
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Nucleic Acids Research, Vol. 18, No. 5
DNA sequence of exoenzyme 03, an ADPribosyltransferase encoded by Clostridium botulinum and D phages
C
Michel Popoff, Patrice Boquet1, D.Michael Gill2* and Melvin W.Eklund3 Unite des Anaerobies and 1Unite des Antigenes Bacteriens, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris, France, 2Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111 and 3Northwest and Alaska Fisheries Center, 2725 Montlake Boulevard East, Seattle, WA 98112, USA Submitted January 29, 1990
EMBL accession no. X51464
Exoenzyme C3 ADP-ribosylates the rho group of small GTPbinding proteins (1, 2). Extracellular C3 from C. botulinum type D is a basic protein of approximate size 26,000 and N terminal AYSNTYQEFTNIDQAKAXGNAQYKKY. Oligonucleotides representing this sequence hybridize to C and D botulinal phage DNA. The nucleotide sequence below was derived from cloned fragments of D phage and represents the extracellular protein of 211 amino acid residues Mr 23,546) plus seven N-terminal amino acid residues which imply a precursor protein cleaved at the site shown with an asterisk. The adjacent 5' DNA sequence is unstable in E. coli and has not been cloned. A candidate transcription terminator (AG = -16 kcalorie) is underlined. C phage have the identical coding sequence and two differences
in the 3' untranslated region: A662, T736. The C3 gene is distinct from the C1 or D neurotoxin genes which are also encoded by these phages (3, 4).
REFERENCES 1. Chardin,P., Boquet,P., Madaule,P., Popoff,M.R., Rubin,E.J. and Gill,D.M. (1989) EMBO J. 8, 1087 - 1092. 2. Rubin,E.J., Gil,D.M., Boquet,P. and Popoff,M.R. (1988) Mol. Cell. Biol. 8, 418-426. 3. Eklund,M.W., Poysky,F.T., Reed,S.M. and Smith,C.A. (1971) Science 172, 480-481. 4. Eklund,M.W., Poysky,F.T. and Reed,S.M. (19972) Nature New Biology 235, 16-17.
*
GCT TAT TCC Ala Tyr Ser CM GCA AAA Gln Ala Lys GM GCT ATA Glu Ala Ile MG GGA GTT
Lys Gly Val TCT TTT AAT Ser Phe Asn TAT Tyr GCT Ala AGT Ser
TTA Leu TTT Phe ACT
GGA
Gly GM Glu TCA
Thr Ser
GTA GCA AAA
Val Ala Lys GAA ATG TTG Glu Met Leu GGT AAA CM Gly Lys Gln AGC TAT TAG AAG TAG CTC
*
ATT AAT CAA AAG GCT TAT TCA MT ACT TAC CAG GAG TTT ACT MT ATT CAT Ile Asn Gln Lys Ala Tyr Ser Asn Thr Tyr Gln Glu Phe Thr Asn Ile Asp GCT 'TGG GGT MT GCT CAG TAT AAA MG TAT GGA CTA AGC AAA TCA GAA AAA Ala 'Trp Gly Asn Ala Gln Tyr Lys Lys Tyr Gly Leu Ser Lys Ser Glu Lys GTA 'TCA TAT ACT AM AGC GCT AGT GM ATA AAT GGA AAG CTA AGA CAA MT Val 'Ser Tyr Thr Lys Ser Ala Ser Glu Ile Asn Gly Lys Leu Arg Gln Asn ATC IMAT GGA TTT CCT TCA AAT TTA ATA AAA CAA GTT GAA CTT TTA GAT AAA Ile IAsn Gly Phe Pro Ser Asn Leu Ile Lys Gln Val Glu Leu Leu Asp Lys AAA iATG AAG ACC CCT GM MT ATT ATG TTA TTT AGA GGC GAC GAC CCT GCT Lys IMet Lys Thr Pro Glu Asn Ile Met Leu Phe Arg Gly Asp Asp Pro Ala ACA (GM TTT CM MC ACT CTT CTT MT TCA AAT GGT ACA ATT AAT AM ACG Thr (Glu Phe Gln Asn Thr Leu Leu Asn Ser Asn Gly Thr Ile Asn Lys Thr MG (GCT AAA GCT MG TTT TTA MT AAA GAT AGA CTT GM TAT GGA TAT ATT Lys AAla Lys Ala Lys Phe Leu Asn Lys Asp Arg Leu Glu Tyr Gly Tyr Ile TTA jATG MT GTT TCT CM TTT GCA GGA AGA CCA ATT ATT ACA AAA TTT AAA Leu IMet Asn Val Ser Gln Phe Ala Gly Arg Pro Ile Ile Thr Lys Phe Lys GGC TCA AAG GCA GGA TAT ATT GAC CCT ATT AGT GCT TTT GCA GGA CM CTT Gly Ser Lys Ala Gly Tyr Ile Asp Pro Ile Ser Ala Phe Ala Gly Gln Leu CTT CCT AGA CAT AGT ACT TAT CAT ATA GAC GAT ATG AGA TTG TCT TCT GAT Leu Pro Arg His Ser Thr Tyr His Ile Asp Asp Met Arg Leu Ser Ser Asp ATA ATA ATT ACA GCA ACA ATG ATG GGC ACA GCT ATC MT CCT AAA TM TAG Ile Ile Ile Thr Ala Thr Met Met Gly Thr Ala Ile Asn Pro Lys *** *** CAT GTT MG GM TTG TAT ATA ATT AM TGT MA MG AGT TAC TTT ATA ACA TTT TTA CTA TM GCA AAA ATA AGA CTG CTA TTT ATA CCC MT ATA TGG CM
To whom correspondence should be addressed
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720 780
