6716 Nucleic Acids Research VoL 18, No. 22
.z) 1990 Oxford University Press
Yeast mitochondrial DNA polymerase is related to the family A DNA polymerases Junetsu Ito and Dan K.Braithwaite Department of Microbiology and Immunology, College of Medicine, University of Arizona Health Sciences Center, Tucson, AZ 85724, USA Submitted October 9, 1990
DNA polymerases have been classified, based on their primary sequence similarities, into two major groups: family A and family B (1). Family A DNA polymerases, represented by E. coli DNA polymerase I (2), includes Streptococcus pneumoniae DNA polymerase I (S.p. pol 1)(3), Thermus aquics DNA polymerase I (Taq pol I)(4), and the DNA polymerases of bacteriophages T5 (5), T7 (6), and Spo2 (7,8). All of these DNA polymerases are prokaryotic and are sensitive to dideoxynucleotide inhibitor, but are resistant to aphidicolin. Family B DNA polymerases, represented by eukaryotic replicative DNA polymerases alpha and delta, are sensitive to aphidicolin and relatively resistant to dideoxynucleotides. Here we show that yeast mitochondrial DNA polymerase I (DNA polymerase 'y) is homologous to family A. Shown in amino acid sequence alignment, yeast mitochondrial DNA polymerase is significanty homologous to family A in both the 3'-5' exonuclease and DNA polymerase domains. Though contradictory to the initial report by Foury (9), these observations
3' Nipi E.c. pot I E.c. DNR Q S.p. pot I
15 T7
B. 141 E.c. pot I S.p. pol I Taq Po 1
75 17
are analogous to the homology of yeast mitochondrial RNA polymerase to T3 and T7 RNA polymerases (11). These findings are consistent with the notion that mitochondria evolved from endosymbiotic prokaryotes (12).
REFERENCES 1. 2. 3. 4. 5.
6. 7.
8. 9. 10. 11. 12.
Jung,G. et al. (1987) Proc. Natl. Acad. Sci. USA 84, 8287-8291. Joyce,C. M. et al. (1982) J. Biol. C/en. 257, 1958-1964. Lopez, P. et al. (1989) J. Biol Chem. 264, 4255-4263. Lawyer,F.C. et al (1989) J. Biol. Chem. 264, 6427-6437. Leavitt,M.C. and Ito,J. (1989) Proc. Natl Acad. Sci. USA 86, 4465-4469. Dunn,J.J. and Studier,F.W. (1983) J. Mol. Biol 166, 477-535. Raden,B. and Rutherg,L. (1984) J. Virol. 52, 9-15. Iwabe,N. et al (1989) Nuct Acids Res. 7, 8866. Foury,F. (1989) J. Biol. Chem 264, 20552-20560. Echols,H. et al. (1983) Proc. Natl. Acad. Sci. USA 80, 2189-2192. Masters,B.S. et al. (1987) Cett 51, 89-99. Mereschowsky,C. (1905) Biol. Zentratbl. 25, 593-604.
5' exonuclease domain
I II 167 LVVF D5V ETLYN. . .D QVVIGHNVAYD RARVLEEYN...337 351 VFAF DTETDSL...414 ALKVGQNLKYDRGILANYGI ..491 8 QIVL DITIEITTGM... 93 AELVIHNNAAFDIGFMDYEFS...14 a6% YSGPIDIVlEINLGK. .32 FELFGENYHT3DNLVGFAWSC ...414 134 PVAFIDISIEITSAL ... f HTIVFHNJLKIFDNMHFYKYHLG ... 2D I MIVSjDI[EANAL ... 5 LIVFH[G HKYDVPALTKLAK ..1.1 3 TLSI DI ETFSS ... 68 VIKTAY AN FERTCIAKHFN ... 1.0
III FQKLANYCAT DVTATSQVFD
LEEAGRIYIAAE DIADVTLQLHL DALCARYEIDNSKRTLHGAL
SLYGQTIYILVDIDIETFYGKGVK FDIMWPIYIAAKIDITDATIRLHN NEENMD[YNVQ[DVVVTKALLE WEKFKVYCIQDVEVERAIKN
DNA polymerase domain I II 68B CFVGADVDSEELWIAS....744 SRMEAKIFNYGRIYGAGA.... 700 VIVSADY QIELRIM.. .753 QRRS AKAINFGLIYGMSA ...W8 648 VLLSSDYISIQIELRVLAI...7 DRRNAKAVNFGVVYGISD...0ZS 606 LLVALDYIS IQELRVLAI .65 MRRAAKTINFGVLYGMSA 77 46 RVIAUDLTITAEVYYAIAI. 1 7 LRQAAKAITFGILYGSGP 47 VQAGIDAISIGLELRCLAI ...517 TRDNAKTFIYGFLYGAGD...646 a 31 E FYVSD F A I RV IJ. L R QK GK VAE LGYQGGK. .99 ..
.
.
.
.
...
.
.
.
.
III A
LCfI.$IHDEIRFLVSE
VIR ININQVHDELVFEVHK TIK ILLQVHDEIVLEVPK AIRMLLQVNDELVLEAPK
M1KIVMNLVHDI!VVAIVRE
FAYMIAW VHDEIQVGCRT YgJTV
MHVDEA
VLDVPR
Figure 1. Regions of homology among the family A DNA polymerases: yeast mitochondrial DNA polymerase (Mipl)(9), E. coli pol I (2), E. coli DNA Q (10), S.p. pol 1 (3), T5 (5), T7 (6), Spo2 (7), Taq pol I (4).
.z) 1990 Oxford University Press
Yeast mitochondrial DNA polymerase is related to the family A DNA polymerases Junetsu Ito and Dan K.Braithwaite Department of Microbiology and Immunology, College of Medicine, University of Arizona Health Sciences Center, Tucson, AZ 85724, USA Submitted October 9, 1990
DNA polymerases have been classified, based on their primary sequence similarities, into two major groups: family A and family B (1). Family A DNA polymerases, represented by E. coli DNA polymerase I (2), includes Streptococcus pneumoniae DNA polymerase I (S.p. pol 1)(3), Thermus aquics DNA polymerase I (Taq pol I)(4), and the DNA polymerases of bacteriophages T5 (5), T7 (6), and Spo2 (7,8). All of these DNA polymerases are prokaryotic and are sensitive to dideoxynucleotide inhibitor, but are resistant to aphidicolin. Family B DNA polymerases, represented by eukaryotic replicative DNA polymerases alpha and delta, are sensitive to aphidicolin and relatively resistant to dideoxynucleotides. Here we show that yeast mitochondrial DNA polymerase I (DNA polymerase 'y) is homologous to family A. Shown in amino acid sequence alignment, yeast mitochondrial DNA polymerase is significanty homologous to family A in both the 3'-5' exonuclease and DNA polymerase domains. Though contradictory to the initial report by Foury (9), these observations
3' Nipi E.c. pot I E.c. DNR Q S.p. pot I
15 T7
B. 141 E.c. pot I S.p. pol I Taq Po 1
75 17
are analogous to the homology of yeast mitochondrial RNA polymerase to T3 and T7 RNA polymerases (11). These findings are consistent with the notion that mitochondria evolved from endosymbiotic prokaryotes (12).
REFERENCES 1. 2. 3. 4. 5.
6. 7.
8. 9. 10. 11. 12.
Jung,G. et al. (1987) Proc. Natl. Acad. Sci. USA 84, 8287-8291. Joyce,C. M. et al. (1982) J. Biol. C/en. 257, 1958-1964. Lopez, P. et al. (1989) J. Biol Chem. 264, 4255-4263. Lawyer,F.C. et al (1989) J. Biol. Chem. 264, 6427-6437. Leavitt,M.C. and Ito,J. (1989) Proc. Natl Acad. Sci. USA 86, 4465-4469. Dunn,J.J. and Studier,F.W. (1983) J. Mol. Biol 166, 477-535. Raden,B. and Rutherg,L. (1984) J. Virol. 52, 9-15. Iwabe,N. et al (1989) Nuct Acids Res. 7, 8866. Foury,F. (1989) J. Biol. Chem 264, 20552-20560. Echols,H. et al. (1983) Proc. Natl. Acad. Sci. USA 80, 2189-2192. Masters,B.S. et al. (1987) Cett 51, 89-99. Mereschowsky,C. (1905) Biol. Zentratbl. 25, 593-604.
5' exonuclease domain
I II 167 LVVF D5V ETLYN. . .D QVVIGHNVAYD RARVLEEYN...337 351 VFAF DTETDSL...414 ALKVGQNLKYDRGILANYGI ..491 8 QIVL DITIEITTGM... 93 AELVIHNNAAFDIGFMDYEFS...14 a6% YSGPIDIVlEINLGK. .32 FELFGENYHT3DNLVGFAWSC ...414 134 PVAFIDISIEITSAL ... f HTIVFHNJLKIFDNMHFYKYHLG ... 2D I MIVSjDI[EANAL ... 5 LIVFH[G HKYDVPALTKLAK ..1.1 3 TLSI DI ETFSS ... 68 VIKTAY AN FERTCIAKHFN ... 1.0
III FQKLANYCAT DVTATSQVFD
LEEAGRIYIAAE DIADVTLQLHL DALCARYEIDNSKRTLHGAL
SLYGQTIYILVDIDIETFYGKGVK FDIMWPIYIAAKIDITDATIRLHN NEENMD[YNVQ[DVVVTKALLE WEKFKVYCIQDVEVERAIKN
DNA polymerase domain I II 68B CFVGADVDSEELWIAS....744 SRMEAKIFNYGRIYGAGA.... 700 VIVSADY QIELRIM.. .753 QRRS AKAINFGLIYGMSA ...W8 648 VLLSSDYISIQIELRVLAI...7 DRRNAKAVNFGVVYGISD...0ZS 606 LLVALDYIS IQELRVLAI .65 MRRAAKTINFGVLYGMSA 77 46 RVIAUDLTITAEVYYAIAI. 1 7 LRQAAKAITFGILYGSGP 47 VQAGIDAISIGLELRCLAI ...517 TRDNAKTFIYGFLYGAGD...646 a 31 E FYVSD F A I RV IJ. L R QK GK VAE LGYQGGK. .99 ..
.
.
.
.
...
.
.
.
.
III A
LCfI.$IHDEIRFLVSE
VIR ININQVHDELVFEVHK TIK ILLQVHDEIVLEVPK AIRMLLQVNDELVLEAPK
M1KIVMNLVHDI!VVAIVRE
FAYMIAW VHDEIQVGCRT YgJTV
MHVDEA
VLDVPR
Figure 1. Regions of homology among the family A DNA polymerases: yeast mitochondrial DNA polymerase (Mipl)(9), E. coli pol I (2), E. coli DNA Q (10), S.p. pol 1 (3), T5 (5), T7 (6), Spo2 (7), Taq pol I (4).
