Nucleic Acids Research, Vol. 18, No. 17 5309
1990 Oxford University Press
Nucleotide sequence of rat hsp60 (chaperonin, GroEL homolog) cDNA Thomas J.Venner and Radhey S.Gupta* Department of Biochemistry, McMaster University, Hamilton, Ontario L8N 3Z5, Canada EMBL accession
Submitted June 26, 1990
The 60 kDa heat shock family of protein (hsp 60) comprise major antigens of pathogenic bacteria (e.g. Mycobacterium tuberculosis, M. leprae) (1). Autoimmune response to hsp 60 has been implicated in a number of autoimmune diseases (e.g. rheumatoid arthritis) (2). This report describes the cloning and sequencing of rat hsp60 cDNA. A Xgtl 1 cDNA library from rat kidney was screened using cDNA probes derived from Chinese hamster hsp60 cDNA (3). For the clones obtained both DNA strands were sequenced using oligonucleotide primers. The clones obtained lacked the 5'- leader sequence as well as the mitochondrial targeting sequence. However, the entire coding sequence for the mature hsp60 protein (amino acids 1 -547) could be deduced. 1
181
271
361
451
541
631
721
901
991
1081
1171
1261
1351 1441
1531
1621 1732 1850
REFERENCES
2. Van Eden,W. et al. (1989) Rheumatoll. 9, 187-191. 3. Picketts,D.J. al. (1989) J. Biol. Chem. 264, 12001-12008. 4. Jindal,S. et al. (1989) Mol. Cell Biol. 9, 2279-2283. et
GCC AAA GTGTA AAATTT
O G GATCCG GTTA ATCTT CM GTTA GAC CTT TTA OCC GAT GCT GTA GCT TT ACAATG GGG t A L N L Q C V D L L AD A V A V T UG K D V K F C A DA AAG OG AMA ACA TO ATT ATT GC CM MT TOG OGA MT = MA PA ACA AM OAT TCA ATT OAT "A GTC ACT PT GCA CS P K V T K D C V T V K S I D T V I I E QS C P KG A L G AM GAT AM TAC AM MT ATC GGA OCT AM CTT PT CM GAT PT GCC MT MC ACA MT MG OCT 00G GAT GGCC MC ACT CCC D E E T T T A N T K D K Y K N I G A K L V Q D V A A ACT GTT CTG GCA CGCTCT AT 0CC AM ATC COG GM GC TTT CMG A ATC AGC AM GOG OCT MT CCA GTG GA OGT GTG ATG TTG P V E I L R R G V T V L A R S I A K E C F E K I S K G A CTT AM AM CM TCT AM CCT GCT GTT GAT OCT PTA ATT OCT ATT OCT CM OTT OCT MA ATT TCT OCA TG MCA CC CCT S V T I A V D A V I A E L K K Q S K P V T T P E E I A A A MC GGA GAC AM GC ATT G0 AMC ATC AT TCT OAT CA ATG A GA G T GGA MA AMG GT GTC ATC ACA OT AMG OAT GGA A MCC S D K T I I A N G D K D I C M K K V G R K C V I T V K D CTO MT GAT GAG CTA GOC ATG AM TTT OAT MA GOA TAT ATT TCC CCA TAT m ATT MC MA TCA AM AT AU GGT CM AM C S C I S P T K K Y F I T L N D E L E I I E C N K F D 0 TCM AM A ATT TCT ATGTT CAG TCC ATT TGT GM TTC CMOAT GCC TAT GTT TTG TT ACCT CT CTT ATT GCCMT GCT C E F Q D A T V L L S E K K I S S V 0 S I V P A L E I A N A CAC CGG MG A CT TTG TC ATA ATT GCT CAT T AT GA CM GCTCTTA GCA CTG GTT TTC GT OGT CTT CAG MAC TG E L S T L V L L K V L A G N R K P L V I I A E D V D GTT GTA GCA GTC AMA OCT CCA 0G0 TTT GCM MC MAGG AM MC CAG CTT AM OAT ATG OCT ATC OCT ACT GGT GGT GCC GTG TTT GGA T V F Q L K D N A I G GA V V A V K A P C F C D N t K GMAGA GGT TTG MT CTAMT CTTCMA T TT C OCTCAT GAT TTA GOG AGTT GGA AG TC ATC GTC ACC AAAT AT CC ATG C E D V L D L K V E V I V K D D N L 0 L E A G T E A CTT TTG AA AM CT ATT CM ATC ACT CMG GGA AM 0GT CM AM GCT CAC ATT TAT GAO CMG CTA OAC ATC MA MT MT E I T E Q L D I T T S E Y E L L K G K G D K A N I E K R I MAG GAMMGCMCA GGCACTT GCT ACTC TCA AT GA GTA GCT GTG TTG MG GTT GGA GGG AAGTOTOT GTC MG TGA A C T S D V E V E K E K L N E R L A K L S D G V A V L K V MG AM GAC GA GTT MA OAT GCT CTC MT GCT MA MA GCA OCT GTT GGC ATT OTT CTA GA 0G0 CGC TOT OCT CTA CTT CGG V T D A L N A T R A A V E E C I V L C G G C A L L K K D MT G T CMG TGC ATC CCA GCC TTG GAT TCA TTA MG C GCA CTT AATT CCT GCA MG ATA G ATA GM ATT ATTAA I I E Q K I I K D E G A L D S L K P A P C I R A L K I P A ATG ACA ATT GCT MG AT GCA GGT GTT CMAG TCT TTG ATA GTT CA MA T CTG CAG AGTTCC TCA GAG GTT GOC TAT GAT GCC ATG V E C S L I V E K I L Q S S S E V C Y D N A C T I A K CTT G A AT TT GTGTMC ATG GTG CMMAGGA ATC ATT GAT CCA ACA MG GTT GTA AACT GCT TTA CTG ATCT GCT GGGGTG CC I D P T K V V T A L L D A A C V A V E K C I D F V N L TCC CTG CTA ACT ACA GCC CG ATT CCT MGACC GCT GTA GTG ACA GA ccAAGC CCT GAATGGT GCAATGOGT GA ATG GGA GGG S L L T T A E A V V T E I P K E E K D PG NG A UG GON GGT ATG GGA GGT GGC ATG UC TM CTCCTAGMTATOCTTTGCCCTTATCATGMCTGTGGCAGGTGCTCAGGCAGGTTCCTCACCAATMCTTCAGAGATCACC C F * 547 N
1969
*
To whom
30
CC
TGMGAMATGACTGCAMAGAGCTCOCTGATCACTGTMCCATCAGTTACTGGTTTCCTTTGACAATATMTATTTTACTOCTGTCATTGTCCATOCCTACAGATMATTTATTTT CTGGGGCGCCTGTCCTGCTTTCAACTTACTOAGCATCTCTACTGTTCTCTTTAGCC
GTATTTTTGATAAAGACATTTGTACATCCT TGTGTCACCACATGAGAGTT
correspondence
should be addressed
X53585
The deduced sequence of rat hsp60 protein differed from the corresponding proteins from human and Chinese hamster cells in 13 and 6 amino acids, respectively, most of which are conservative replacements (3, 4).
A
91
no.
TCTGTGGAGGGTAGATGATTGTGTACAGAGTAGAGAGTATCCATTATGTGACACCTTTGTGTMTAATTTTGTTTA
60 90
120 150
180
210 240
270 300
330
360 390 420
450
40 510 540
1990 Oxford University Press
Nucleotide sequence of rat hsp60 (chaperonin, GroEL homolog) cDNA Thomas J.Venner and Radhey S.Gupta* Department of Biochemistry, McMaster University, Hamilton, Ontario L8N 3Z5, Canada EMBL accession
Submitted June 26, 1990
The 60 kDa heat shock family of protein (hsp 60) comprise major antigens of pathogenic bacteria (e.g. Mycobacterium tuberculosis, M. leprae) (1). Autoimmune response to hsp 60 has been implicated in a number of autoimmune diseases (e.g. rheumatoid arthritis) (2). This report describes the cloning and sequencing of rat hsp60 cDNA. A Xgtl 1 cDNA library from rat kidney was screened using cDNA probes derived from Chinese hamster hsp60 cDNA (3). For the clones obtained both DNA strands were sequenced using oligonucleotide primers. The clones obtained lacked the 5'- leader sequence as well as the mitochondrial targeting sequence. However, the entire coding sequence for the mature hsp60 protein (amino acids 1 -547) could be deduced. 1
181
271
361
451
541
631
721
901
991
1081
1171
1261
1351 1441
1531
1621 1732 1850
REFERENCES
2. Van Eden,W. et al. (1989) Rheumatoll. 9, 187-191. 3. Picketts,D.J. al. (1989) J. Biol. Chem. 264, 12001-12008. 4. Jindal,S. et al. (1989) Mol. Cell Biol. 9, 2279-2283. et
GCC AAA GTGTA AAATTT
O G GATCCG GTTA ATCTT CM GTTA GAC CTT TTA OCC GAT GCT GTA GCT TT ACAATG GGG t A L N L Q C V D L L AD A V A V T UG K D V K F C A DA AAG OG AMA ACA TO ATT ATT GC CM MT TOG OGA MT = MA PA ACA AM OAT TCA ATT OAT "A GTC ACT PT GCA CS P K V T K D C V T V K S I D T V I I E QS C P KG A L G AM GAT AM TAC AM MT ATC GGA OCT AM CTT PT CM GAT PT GCC MT MC ACA MT MG OCT 00G GAT GGCC MC ACT CCC D E E T T T A N T K D K Y K N I G A K L V Q D V A A ACT GTT CTG GCA CGCTCT AT 0CC AM ATC COG GM GC TTT CMG A ATC AGC AM GOG OCT MT CCA GTG GA OGT GTG ATG TTG P V E I L R R G V T V L A R S I A K E C F E K I S K G A CTT AM AM CM TCT AM CCT GCT GTT GAT OCT PTA ATT OCT ATT OCT CM OTT OCT MA ATT TCT OCA TG MCA CC CCT S V T I A V D A V I A E L K K Q S K P V T T P E E I A A A MC GGA GAC AM GC ATT G0 AMC ATC AT TCT OAT CA ATG A GA G T GGA MA AMG GT GTC ATC ACA OT AMG OAT GGA A MCC S D K T I I A N G D K D I C M K K V G R K C V I T V K D CTO MT GAT GAG CTA GOC ATG AM TTT OAT MA GOA TAT ATT TCC CCA TAT m ATT MC MA TCA AM AT AU GGT CM AM C S C I S P T K K Y F I T L N D E L E I I E C N K F D 0 TCM AM A ATT TCT ATGTT CAG TCC ATT TGT GM TTC CMOAT GCC TAT GTT TTG TT ACCT CT CTT ATT GCCMT GCT C E F Q D A T V L L S E K K I S S V 0 S I V P A L E I A N A CAC CGG MG A CT TTG TC ATA ATT GCT CAT T AT GA CM GCTCTTA GCA CTG GTT TTC GT OGT CTT CAG MAC TG E L S T L V L L K V L A G N R K P L V I I A E D V D GTT GTA GCA GTC AMA OCT CCA 0G0 TTT GCM MC MAGG AM MC CAG CTT AM OAT ATG OCT ATC OCT ACT GGT GGT GCC GTG TTT GGA T V F Q L K D N A I G GA V V A V K A P C F C D N t K GMAGA GGT TTG MT CTAMT CTTCMA T TT C OCTCAT GAT TTA GOG AGTT GGA AG TC ATC GTC ACC AAAT AT CC ATG C E D V L D L K V E V I V K D D N L 0 L E A G T E A CTT TTG AA AM CT ATT CM ATC ACT CMG GGA AM 0GT CM AM GCT CAC ATT TAT GAO CMG CTA OAC ATC MA MT MT E I T E Q L D I T T S E Y E L L K G K G D K A N I E K R I MAG GAMMGCMCA GGCACTT GCT ACTC TCA AT GA GTA GCT GTG TTG MG GTT GGA GGG AAGTOTOT GTC MG TGA A C T S D V E V E K E K L N E R L A K L S D G V A V L K V MG AM GAC GA GTT MA OAT GCT CTC MT GCT MA MA GCA OCT GTT GGC ATT OTT CTA GA 0G0 CGC TOT OCT CTA CTT CGG V T D A L N A T R A A V E E C I V L C G G C A L L K K D MT G T CMG TGC ATC CCA GCC TTG GAT TCA TTA MG C GCA CTT AATT CCT GCA MG ATA G ATA GM ATT ATTAA I I E Q K I I K D E G A L D S L K P A P C I R A L K I P A ATG ACA ATT GCT MG AT GCA GGT GTT CMAG TCT TTG ATA GTT CA MA T CTG CAG AGTTCC TCA GAG GTT GOC TAT GAT GCC ATG V E C S L I V E K I L Q S S S E V C Y D N A C T I A K CTT G A AT TT GTGTMC ATG GTG CMMAGGA ATC ATT GAT CCA ACA MG GTT GTA AACT GCT TTA CTG ATCT GCT GGGGTG CC I D P T K V V T A L L D A A C V A V E K C I D F V N L TCC CTG CTA ACT ACA GCC CG ATT CCT MGACC GCT GTA GTG ACA GA ccAAGC CCT GAATGGT GCAATGOGT GA ATG GGA GGG S L L T T A E A V V T E I P K E E K D PG NG A UG GON GGT ATG GGA GGT GGC ATG UC TM CTCCTAGMTATOCTTTGCCCTTATCATGMCTGTGGCAGGTGCTCAGGCAGGTTCCTCACCAATMCTTCAGAGATCACC C F * 547 N
1969
*
To whom
30
CC
TGMGAMATGACTGCAMAGAGCTCOCTGATCACTGTMCCATCAGTTACTGGTTTCCTTTGACAATATMTATTTTACTOCTGTCATTGTCCATOCCTACAGATMATTTATTTT CTGGGGCGCCTGTCCTGCTTTCAACTTACTOAGCATCTCTACTGTTCTCTTTAGCC
GTATTTTTGATAAAGACATTTGTACATCCT TGTGTCACCACATGAGAGTT
correspondence
should be addressed
X53585
The deduced sequence of rat hsp60 protein differed from the corresponding proteins from human and Chinese hamster cells in 13 and 6 amino acids, respectively, most of which are conservative replacements (3, 4).
A
91
no.
TCTGTGGAGGGTAGATGATTGTGTACAGAGTAGAGAGTATCCATTATGTGACACCTTTGTGTMTAATTTTGTTTA
60 90
120 150
180
210 240
270 300
330
360 390 420
450
40 510 540
