Redox regulation of SurR by protein disulfide oxidoreductase in ThermococcusonnurineusNA1
Jae KyuLim1,2·Hae-Chang Jung1,2·Sung GyunKang1,2·Hyun Sook Lee1,2
Sung Gyun Kang · Hyun Sook Lee
e-mail:,
1 Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan15627, Republic of Korea
2 Department of Marine Biotechnology, Korea University of Science and Technology, Daejeon 34113, Republic of Korea
Running title: Reduction of SurR by protein disulfide oxidoreductase
Supplementary material accompanies this paper.
Table S1Sequences of the oligonucleotide primers used for PCR amplification.
Primer / Location / Oligonucleotide sequenceGene cloning
prnTON0470F / TON_0470 forward / 5'- TGCCGCGCGGCAGCCATATGAGGAGGACTGCTTTACTGTTG-3'
prnTON0470R / TON_0470 reverse / 5'- TGGTGGTGGTGGTGCTCGAGGCGCCTTCTCCTCAGGAG-3'
prnTON0472F / TON_0472 forward / 5'- TGCCGCGCGGCAGCCATATGGACGAGCTGGAGATGATC-3'
prnTON0472R / TON_0472 reverse / 5'- TGGTGGTGGTGGTGCTCGAGCTGTAGGCCTAGCAGCATCC-3'
prnTON0834F / TON_0834 forward / 5'- TGCCGCGCGGCAGCCATATGATCAGGGAGTTTGATGGAG-3'
prnTON0834R / TON_0834 reverse / 5'- TGGTGGTGGTGGTGCTCGAGGGAGTCAGAACCCAGGAGC-3'
prnTON0318F / TON_0318 forward / 5'- TGCCGCGCGGCAGCCATATGACTGAACCGGACATCTTTTAC-3'
prnTON0318R / TON_0318 reverse / 5'- TGGTGGTGGTGGTGCTCGAGATCTTCTCCGGCATCTTCGAC-3'
EMSA DNA probe
prnPmbhF / mbh promoter forward / 5'-CATCCAATTAGGTTTGGGAAATGGT-3'
prnPmbhR / mbh promoter reverse / 5'-AAGGCGGCAATAAATGCCATCTTTC-3'
1
Table S2 Conservation of genes encoding Trx- or Grx-like proteins in the genomes of the order Thermococcales
Genome / Homologues of T. onnurineus NA1 protein (% identity)TON_0319 / TON_0470 / TON_0472 / TON_0834
Thermococcalesarchaeon44_46 / XD43_0781 (77.7) / XD43_1009 (39.5) / XD43_1823 (80.9) / - b
ThermococcusbarophilusMP / TERMP_00657 (81.7) / TERMP_RS04160 (38.8) / TERMP_00838 (76.7) / TERMP_01376 (51.5)
Thermococcuscelericrescens / APY94_12140 (91.6) / APY94_05295 (60.6) / APY94_05305 (85.2) / APY94_01785 (49.5)
Thermococcuscleftensis / CL1_0385 (93.8) / CL1_0213 (63.1) / CL1_0211 (87.9) / -
Thermococcuseurythermalis / TEU_06555 (90.3) / TEU_08320 (48.2) / - / -
ThermococcusgammatoleransEJ3 / TGAM_1302 (89.8) / TGAM_1636 (42.8) / - / -
ThermococcusguaymasensisDSM 11113 / X802_10250 (84.1) / X802_01105 (46.0) / - / -
ThermococcuskodakarensisKOD1 / TK1085 (88.5) / TK1208 (53.3) / - / TK0838 (50.5)
ThermococcuslitoralisDSM 5473 / OCC_08455 (78.1) / OCC_10925 (40.8) / OCC_13031 (79.1) / OCC_05726 (53.5)
Thermococcus nautili / BD01_0185 (87.2) / BD01_0376 (47.7) / BD01_0378 (88.7) / -
Thermococcusparalvinellae / TES1_0610 (83.0) / TES1_0848 (40.4) / TES1_0851 (77.6) / TES1_1390 (87.4)
Thermococcuspeptonophilus / A0127_09800 (88.5) / A0127_08885 (48.5) / A0127_00965 (83.5) / A0127_10085 (53.5)
Thermococcussibiricus / TSIB_0991 (72.8) / TSIB_RS09715 (37.4) / TSIB_1746 (77.4) / TSIB_1703 (50.5)
Thermococcussp. 4557 / GQS_03675 (91.2) / GQS_02835 (51.9) / - / -
Thermococcusthioreducens / AMR53_03845 (91.6) / AMR53_01730 (60.3) / AMR53_01740 (85.2) / AMR53_05300 (50.0)
Thermococcuszilligii / WP_010478330a (80.4) / WP_010478037a(49.2) / WP_010479774a(83.5) / -
PyrococcusabyssiGE5 / PAB2245 (83.5) / PAB1651 (30.3) / - / -
Pyrococcus furiosus DSM 3638 / PF0094 (80.5) / PF0724 (43.2) / PF1457 (80.7) / PF0741 (47.0)
PyrococcushorikoshiiOT3 / PH0178 (82.1) / PH1130 (34.0) / - / -
Pyrococcus sp. NA2 / PNA2_0698 (83.6) / PNA2_1761 (36.0) / - / -
Pyrococcussp. ST04 / Py04_0249 (84.5) / Py04_1135 (36.1) / - / Py04_0921 (48.5)
PyrococcusyayanosiiCH1 / PYCH_16300 (88.0) / PYCH_08480 (43.6) / PYCH_08820 (80.9) / PYCH_09150 (50.5)
Palaeococcusferrophilus / WP_048148193a(80.4) / WP_048150379a(50.2) / WP_048147785a(78.3) / WP_048147736a(50.5)
PalaeococcuspacificusDY20341 / PAP_07225 (80.4) / PAP_09015 (55.5) / PAP_08980 (76.1) / PAP_08695 (78.6)
a NCBI ( reference sequence accessionnumber was used instead of Locus tag.
b- means no detection
1
Fig.S1 Phylogenetic tree of Trx- or Grx-like proteins. The tree was constructed using the Mega 7 program with the neighbor-joining method with 500 bootstrap replicates. Species name along with the locus tag or NCBI reference sequence accession number are indicated at the termination of each branch.
A. TON_0319 homologue
PAP_07225 --MGLISDGDKKVIREEFFSKLTNPVKIIVFIGKDHCQYCEQLKQLVQEVSELSDLVSYE58
TERMP_00657 --MGLISDGDKKVIREEFFSKLTNPVKLIVFIGKDHCQYCEQLKQLVQEISELSDLVSFE58
TES1_0610 --MGLISDGDKKVIKEEFFSKLTNPVKLIVFVGKEHCQYCEQLKQLVQEISELSDLVSYE58
TSIB_0991 --MALISDGDKKIIKEEFFSKLVNPVKLLVFIGKDHCQYCDQLKQLVEEISELSDLVTYE58
XD43_0781 --MALISDGDKKIIKEEFFSKLVNPVKLIVFTGKEHCQYCDQLKQLVEEISELSDLISYE58
OCC_08455 MEMALISDGDKKIIKEEFFSKLVNPVKIIVFTGKEHCQYCDQLKQLVEEISELSDLISYE60
WP_048148193 --MGLISDGDKKVIKEEFFSKLEEPVKLIAFIGKEHCQYCDQLKQILAEISELSDKVTYE58
WP_010478330 --MGLISDADKRVIKEEFFSRLTNPVKLMVFTGREHCQYCTQLKQLVQELSELTDKLTYE58
PF0094 --MGLISDADKKVIKEEFFSKMVNPVKLIVFVRKDHCQYCDQLKQLVQELSELTDKLSYE58
Py04_0249 --MGLISDADKKIIKEEFFSKMTNPVKLIVFIGKEHCQYCDQLKQLVQELSELSDKLTYE58
PNA2_0698 --MGLISEEDKRIIKEEFFSKMVNPVKLIVFIGKEHCQYCDQLKQLVQELSELTDKLSYE58
PAB2245 --MGLISEEDKRIIKEEFFSKMVNPVKLIVFIGKEHCQYCDQLKQLVQELSELTDKLSYE58
PH0178 --MGLISEEDKRIIKEEFFSKMVNPVKLIVFIGKEHCQYCDQLKQLVQELSELTDKLSYE58
PYCH_16300 --MGLISEEDKKVIREEFFSKMTNPVKLIVFVGKEHCQYCDQLKQLVQELSELTDKLTYE58
X802_10250 --MGLISDADKKVIKEEFFSKLTEPVKLMVFTGKEHCQYCDQLKQLVQELAELSDKLTYE58
BD01_0185 --MGLISDADKKVIKEEFFSKLTEPVKLMVFIGKDHCQYCDQLKQLVQELAELSDKLSYE58
TK1085 --MGLISDADKKVIKEEFFSKMTNPVKLIVFVGKEHCQYCDQLKQLVQELSELTDKLSYE58
A0127_09800 --MGLISDADKKVIKEEFFSKMTNPVKLIVFVGKEHCQYCDQLKQLVQELSELSDKLSYE58
AMR53_03845 --MGLISDADKKVIREEFFSKMTNPVKIIGFIGKEHCQYCDQLKQLVQELADLSDKLTYE58
TEU_06555 --MGLISDADKKVIKEEFFSKLTNPVKIIGFIGKDHCQYCDQLKQVVQELSELTDKLTYE58
APY94_12140 --MGLISDADKKVIREEFFSKMTNPVKIIGFIGKEHCQYCDQLKGLVQELSELSDKLTYE58
GQS_03675 --MGLISDADKKVIKEEFFSKMTNPVKIIGFTGKEHCQYCDQLKGLVQELSELTDKLTYE58
TGAM_1302 --MGLISDADKKIIKEEFFSKMTNPVKIIGFTGKEHCQYCDQLKQLVQELSELTDKLTYE58
CL1_0385 --MGLISDNDKKVIKEEFFSKLTNPVKIIGFIGKDHCQYCDQLKQLVQELSELTDKLTYE58
TON_0319 --MGLISDGDKKVIREEFFSKMTNPVKIIGFIGKDHCQYCDQLKQLVQELSELSDKLSYE58
*.***: **::*:*****:: :***:: * ::***** *** :: *:::*:* :::*
PAP_07225 IHDFDNE--KELAEKYRIDKAPSLVITQDGKDLGVRYFGLPAGHEFGAFLEDIVDVSNGT116
TERMP_00657 VHDFDTE--KELAQRYRVDRAPVTVITQDGKDFGVRYFGLPAGHEFGAFLEDIVDVSNGT116
TES1_0610 IHDFDTE--KELAEKYRIDRAPATIITQDGKDLGVRYFGLPAGHEFGAFLEDIVDVSNGT116
TSIB_0991 LHDFDSE--KELAEKYRIDIAPAVVMTQNGKDLGVRYFGLPAGHEFGSFLEDIVDVSKGE116
XD43_0781 IHDFDSE--KELAEKYRVDRAPVTIITQDGKDLGVRYFGLPAGHEFGSFLEDIVDVSRGE116
OCC_08455 VHDFDSE--KELAEKYRIDKAPVTIITQDGKDFGVRYFGLPAGHEFGSFLEDIVDVSKGE118
WP_048148193 VHDFDAE--KELAEKYRVDRAPATVITRNGEDFGVRFFGLPAGHEFGAFLEDIVDVSTGN116
WP_010478330 FHNFDTPEGKEIARKYRIDRAPAVTITQDGKDMGVRYFGLPAGHEFGSFLEDIVDVSNGT118
PF0094 IVDFDTPEGKELAKRYRIDRAPATTITQDGKDFGVRYFGLPAGHEFAAFLEDIVDVSREE118
Py04_0249 IVDFDTPEGQELARKYRIDRAPATTITQDGKDFGVRYFGLPAGHEFAAFLEDIVDVSNAQ118
PNA2_0698 IIDFDTPEGKELAKKYRIDRAPATTITQDGKDFGVRYFGIPAGHEFAAFLEDIVDVSRAD118
PAB2245 IVDFDTPEGKELAEKYRIDRAPATTITQDGKDFGVRYF--PAGHEFAAFLEDIVDVSRAE116
PH0178 IVDFDTPEGKELAEKYRIDRAPATTITQDGKDFGVRYF--PAGHEFAAFLEDIVDVSKGD116
PYCH_16300 VYDFDTEEGRKIAEQYKVDRAPAVTITQDGRDMGVRYFGLPAGHEFAAFLEDIVDVSNAQ118
X802_10250 FIDFDTEEGKRKAEEYRIDHAPAVSITRNGQDVGVRFFGLPAGHEFGAFLEDIVDVSNMT118
BD01_0185 VIDFDTEEGKKKAEEYRIDRAPAVSITRNGQDVGVRFFGLPAGHEFGAFLEDIVDVSNMT118
TK1085 VVDFDTEEGKKLAEQYRIDRAPATTITQDGKDMGVRYFGLPAGHEFGAFLEDIVDVSNGT118
A0127_09800 VVDFDTEEGKKLAEQYRIDRAPATTITQDGRDMGVRFFGLPAGHEFGAFLEDIVDVSNGT118
AMR53_03845 FHDFDSEEGRKLAEQYKIDRAPAVTITQDGRDMGVRFFGLPAGHEFGAFLEDIVDVSNAT118
TEU_06555 FHDFDSEEGRKVAEQYRIDRAPAITITQDGKDMGVRYFGLPAGHEFGALLEDIVDVSNAT118
APY94_12140 FHDFDSEEGRKLAEQYRIDRAPAITLTQDGKDMGVRFFGLPAGHEFGAFLESIVDVSNAT118
GQS_03675 FHDFDTEEGRKIAEQYRIDRAPAITLTQDGKDMGVRFFGLPAGHEFGAFLESIVDVSNAT118
TGAM_1302 FYDFDTEEGRKVAEQYRIDRAPAVTITHDGKDVGVRFFGLPAGHEFSAFLEDIVDVSNMT118
CL1_0385 FHDFDSEEGRKLAEQYRIDRAPAVTITQDGKDMGVRFFGLPAGHEFGAFLEDIVDVSNAS118
TON_0319 FYDFDSEDGRKLAEQYGIDRAPAITITQDGKDMGVRFFGLPAGHEFGAFLEDIVDVSNAQ118
. :** :. *..* :* ** :*::*.*.***:* ******.::**.*****
PAP_07225 TDLMPETKEALKNIDKDVQIYAFVTPTCPYCPMAVRMAHKFAIENALAGKGKILGDMIEA176
TERMP_00657 TDLMKETKETLKEIDRDVRILVFVTPTCPYCPMAVRMAHKFALENALAGKNKILGDMVEA176
TES1_0610 TDLMPDTKDTLKSIDKDVRILVFVTPTCPYCPMAVRMAHKFAIENALAGKGKILGDMVEA176
TSIB_0991 TDLSEETKEALKEINKDVQIYVFVTPTCPYCPMAVRMAHKFAIEHALAGNDRILGDMIEA176
XD43_0781 TDLSEETKEAVRNIDRDVEIYVFVTPTCPYCPMAVRMAHKFAIENALAGKGKILGDMIEA176
OCC_08455 TDLMEDTKEAVRNIDQDVEIYVFVTPTCPYCPMAVRMAHKFAIENSLAGKGKILGDMIEA178
WP_048148193 TDLAPDTKEAIANVERDVRILVFVTPTCPYCPMAVRMAHKFALENTKAGKGKILGDMVEA176
WP_010478330 TDLMPESKKALH--ERDVRILVFVTPTCPYCPLAVRMAHKFAIENAIAGKGRILGDMVEA176
PF0094 TNLMDETKQAIRNIDQDVRILVFVTPTCPYCPLAVRMAHKFAIENTKAGKGKILGDMVEA178
Py04_0249 TDLMDESKEALRNIDQDVRILVFVTPTCPYCPLAVRMAHKFAIENTKAGKGKILGDMVEA178
PNA2_0698 TDLMPDSKEEVAKIDKDVRILVFVTPTCPYCPLAVRMAHKFAIENTKAGKGKILGDMVEA178
PAB2245 TDLMAESKEEVAKIDKNVRILVFVTPTCPYCPLAVRMAHKFAIENTKAGKGKILGDMVEA176
PH0178 TDLMQDSKEEVSKIDKDVRILIFVTPTCPYCPLAVRMAHKFAIENTKAGKGKILGDMVEA176
PYCH_16300 TDLMPDTKETIR--DRDVRILVFVTPTCPYCPMAVRMAHKFAIENTNAGKGKIKGDMVEA176
X802_10250 TDLMPDTKEAIANVDRDVRILVFVTPTCPYCPLAVRMAHKFAVENTKAGKGRILGDMVEA178
BD01_0185 TDLMPESKEALAKIDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGKILGDMVEA178
TK1085 TDLAPDTKEAIHGVDKDVRILVFVTPTCPYCPLAVRMAHKFAIENTLAGKGKILGDMVEA178
A0127_09800 TDLAPDTKEAIHGVDKDVRILVFVTPTCPYCPLAVRMAHKFAIENTLAGKGKILGDMVEA178
AMR53_03845 TDLMPDSKEELAEIDRDVRILVFVTPTCPYCPLAVRMAHKFAIENAKAGKGRILGDMVEA178
TEU_06555 TDLMQETKEAIANVERDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGKILGDMVEA178
APY94_12140 TDLMPESKQDLANVDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGRILGDMVEA178
GQS_03675 TDLMPESKEELAKVDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGKILGDMVEA178
TGAM_1302 TDLMPESKEALAKIDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGRILGDMVEA178
CL1_0385 TDLMPESKEELAKIDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGKILGDMVEA178
TON_0319 TDLMPDTKEELAKVDRDVRILVFVTPTCPYCPLAVRMAHKFAIENTNAGKGKIRGDMVEA178
*:* ::*. : :::*.* **********:*********:*:: **:.:* ***:**
PAP_07225 IEYPELADKYSVMAVPKVVIRVNGEDKVSFEGAYPEKMFLEKLMEALE224
TERMP_00657 IEYPEWADQYRVMAVPKIVIQVDGVDKVEFEGAYPEKMFLEKLLEALE224
TES1_0610 IEYPEWADQYRVMAVPKIVIQVDGVDKVEFEGAYPEKMFLEKLLEALE224
TSIB_0991 IEFPELAEQYGVMAVPKVVIKVNGEDKISFEGAYPEKMFLEKLLEALE224
XD43_0781 IEFPEWADQYSVMAVPKVVIRVDGEDKVSFEGAYPEKMFLEKLLQALE224
OCC_08455 IEFPEWADQYSVMAVPKVVIRVNGEDKVSFEGAYPEKMFLEKLLQALE226
WP_048148193 IEYPEWADEYKVMAVPKVVIMVDGEDKVQFEGAYPEKMFLEKLLAALE224
WP_010478330 IEYPEWADEYSVMAVPKIVIQVDGEDRVEFEGAYPEKMFLEKLLAALE224
PF0094 IEYPEWADQYNVMAVPKIVIQVNGEDRVEFEGAYPEKMFLEKLLSALS226
Py04_0249 IEYPEWADQYNVMAVPKIVIQVNGEDKVQFEGAYPEKMFLEKLLAALS226
PNA2_0698 IEYPEWADQYNVMAVPKIVIQVNGEDKVQFEGAYPEKMFLEKLLAALS226
PAB2245 IEYPEWADQYNVMAVPKIVIQVDGEDKVQFEGAYPEKMFLEKLLAALS224
PH0178 IEYPEWADQYNVMAVPKIVIQVNGEDKVQFEGAYPEKMFLEKLLSALS224
PYCH_16300 MEYPEWADQYNVMAVPKIVIQVDGEDKVQFEGAYPEKMFVEKLLAALG224
X802_10250 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLTALE226
BD01_0185 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLAALE226
TK1085 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFLEKLLAALE226
A0127_09800 IEYPEWADQYSVMAVPKIVIQVDGEDKIQFEGAYPEKMFLEKLLAALE226
AMR53_03845 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLAALS226
TEU_06555 IEYPEWADQYSVMAVPKVVIQVDGEDKVQFEGAYPEKMFLEKLLAALE226
APY94_12140 IEYPEWADKYSVMAVPKIVIIVDGEDKVQFEGAYPEKMFMEKLLAALE226
GQS_03675 IEYPEWADKYSVMAVPKIVIIVDGEDKVQFEGAYPEKMFMEKLLAALE226
TGAM_1302 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLAALE226
CL1_0385 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLAALE226
TON_0319 IEYPEWADQYSVMAVPKIVIQVDGEDKVQFEGAYPEKMFMEKLLAALE226
:*:** *::* ******:** *:* *::.**********:***: **
B. TON_0470 homologue
TSIB_RS09715 ---MKRIVVSLLIM---MMVFISMCISNNTSTSST------29
XD43_1009 ---MKKVLTGLLLV---LVVLTSMCISDNPSSSSS------29
OCC_10925 ---MKKVLTGLLLV---LVVLTSMCISDNPSSSSS------29
TES1_0848 ---MRRILNGILIV-LVLSTVMSMCISNNSSTSTQIASTSNSQATVSTTQVLTTTSTIQS56
TERMP_RS04160 ---MKRIVAGVVFAVVILGVFASLCISNNNTMTSPSAM------Q-36
PYCH_08480 ----MKELSILL--IIILAVAALGCLGETSTPTTSPP------31
Py04_1135 ---MNRVPIILL--VFVLPIVISGCISSPPSTQTISQHAST------36
PH1130 ---MKRDLALLL--ILVLAASFLGCISSQ------24
PAB1651 ----MKKVAFIL--IIVLLAFSLGCLSSTSTPTTTQT--SQ------33
PNA2_1761 ----MRKAFTVI--LLILLTSSLGCLSSN------23
PF0724 ---MKLKAL-FL--LLALGSLIVPRI------20
CL1_0213 ---MRKTGVVLV--LLLIVGVASGCISNSNGPTA---TQT------32
GQS_02835 ---MKKFGIVLL--IILLAGFAAGCISNSGGSNTSSTTQT------35
TK1208 ---MKKTGILLM--LLVLVGFASACLSSDSN--TSS---Q------30
A0127_08885 ---MKKTGILLM--LLVLVGFASACLGSDSN--TSS---Q------30
TEU_08320 ---MKKVGVLLL--LLFLIGFSAGCIGSDNTDTTSS---T------32
BD01_0376 ---MKKAGVLLL--LLVLLGFSAGCIGSNGTTGTSS---T------32
WP_010478037 ---MKVPRALLA--LTLSLAFLALPVASA------24
TGAM_1636 MAGMKRVV-GLA--LLLLLSSVLFPV-SA------25
X802_01105 ---MKRFVMSAT--LLLLVSSMLLPIASA------24
TON_0470 ---MRRTALLLL--FLLTVSLLSYAV------21
APY94_05295 ---MRKLVLIII--GMLLISTVSYAV------21
AMR53_01730 ---MKKLVLFFI--WLLVFSTVSYAL------21
WP_048150379 ---MRKLYGLVI--LLIVLSTLSHAL------21
PAP_09015 ---MRKAALFIV--FLFLASAVSLVL------21
:
TSIB_RS09715 --PTTNPT---ETTNPLTLDKSKFHFYEYGLATCPHCRKMKEELPKAFGKEALTYYELQG84
XD43_1009 --PS------TATETFTLDKAKFHFYMYGLATCPHCKKMKEEIPKFFGKDALTYYELQG80
OCC_10925 --PS------TATETFTLDKAKFHFYMYGLATCPHCKKMKEDIPEFFGEEALTYYELQG80
TES1_0848 SGPSSTET---TSTHEQNIDITKLHFYMFGVSTCPHCGKMKQLLPEIFGKDKFTYYELQG113
TERMP_RS04160 --SSSVNT---QTQSQNQIDVSKLHFYMFGLATCPHCKKMKQLLPEAFGKNSLTYYELQG91
PYCH_08480 ------KLQLDKSKFHFYLYGMATCPHCQNMKRELPKFFGEDSLTYYELIN76
Py04_1135 -TPTSHQSTTITEDWLEGLDKSKFHFYIYGLDTCPHCQRMKELLPEYFGEGSLTFYEVRK95
PH1130 -----TQTQTSQEKWLEGLKKSEFHFYIFGLNTCPHCQRMKKLLPEYFGNSSLTFYEIRE79
PAB1651 -PSPTTPSKTQAQDWLEGLDKSKFHFYVFGVNTCPHCRRMKELLPEYFGNKSLTFYEVRE92
PNA2_1761 -----TPTQTKTESWLEGLEKEKFHFYMFGVNTCPHCQKMKRLLPEYYGNGSLTFYELRE78
PF0724 ------EAEEFESSKVHFYMYGAYWCPHCRNVKESIKEHFGEETLTYYEIEG66
CL1_0213 ---TTSSIPDYVVVNGTKIYLDEIHFYMYGMKTCPHCQNMKKLIPETYGEDSLTYYELVD89
GQS_02835 ---TSSGGSDYVVVNGTKIYLDDIHFYMYGMKTCPHCHKMREEISETYGEDSLTYYELVD92
TK1208 ---TSTESPKYVDVNGTRIYLDQIHFYMYGMKTCPHCRHMKEWIPEEFGADSLTYYELVN87
A0127_08885 ---TSTESPKYVDVNGTKIYLDQIHFYMYGAKTCPHCRNMKTQIPSVYGEDALTYYELID87
TEU_08320 ---STAAQQDYVDVNGTKIYLDKVHFYMYGAKTCPHCRNMKEKIPAKYGEEAFTYYELVN89
BD01_0376 ---STTSKPAYVDVNGTRIYLNEIHFYMYGMKTCPHCRHMHEWIPQEYGEKALTYYELVG89
WP_010478037 ------SDWKFDKSKVHFYMYGLSTCPHCRNMKQLIPEVYGSDKLTYYELNG70
TGAM_1636 ------SDWKFDPSLVHFYLYGAKTCPHCRHMKEVIPETYGADKFTYYELVG71
X802_01105 ------SEWKFDPSLVHFYMYGAKTCPHCRHMKELIPATYGSDKFTYYELTG70
TON_0470 ------AETQIDSNKVHFYMYGMSTCPHCQNMKKLIPEIYGPESLTYYELVN67
APY94_05295 ------AEGQVNPDKLDFYMYGAKTCPHCRKMKEEIPRVYGNDSFTYYELVD67
AMR53_01730 ------VETQIDPGKLHFYMYGLETCPHCQKMKEEIPKFYGENSLTYYELIN67
WP_048150379 ------AQGEIDPKKVHFYMYGAATCPHCKKMKEEIPKAFGADSLTYYELVN67
PAP_09015 ------ADTQIDKSKVHFYMYGTKTCPHCKKMKEEIPKQFGEQSLTYYELID67
. ..** :* **** .:: : :* :*:**:
TSIB_RS09715 NDHNNQMFNQLYEILGVTTVPVIGIFYDGKLYAIVNGEFPTDY--ADDFVEEAMKAKGVL142
XD43_1009 NEHNGKMFEQLYEILGVTGVPVIGVFYEGKLYAIVNGEFPVDY--ADDLVEEAKKAEGVL138
OCC_10925 NEHNGKMFEQLYEILGVTGVPVIGVFYEGKLYAIVNGEFPVDY--ADDLVEEAKKAEGVL138
TES1_0848 NEHNTQLFSELYNILGVTGVPVIGIFYDNHLVAVIEGEIPDPAKNVPWLVNTALKENGVL173
TERMP_RS04160 NDHNSNLFSELYSMLGVTGVPVIGIFYDNRLVAIVEGEIPEPSKNVPIIVETAIKEKGVL151
PYCH_08480 NEHNTKIFMEFSKELGVRGVPLIGVFYDGKLYAIIEGEIDP--SKIPEIVKEAMDREGII134
Py04_1135 SEKNFEIYQNFSKMIGVTGVPLIGVFYSGKLYAIIEGEIDP--KVIPKIVKTAMEHNSVI153
PH1130 DKKAYNTYMKFVKTLGITGVPLIGIFYKDNLYAVVEGEIDP--KVIPQLVKEAMKNNGVI137
PAB1651 NKTAYEFYLKFMETLGVTGVPLIGIFYENKLYAVVEGEIDP--RDIPKLVKAAMKENGVA150
PNA2_1761 NKKAYEFYLNFATTLKITGVPLIGIFYDNKLYAIVEGEIDP--KVIPRIVKEAMKDNGVI136
PF0724 VEHNKNKFEELYLLTGISGIPATAIFYEGEIYAVIEGEFDV--SKTWQIITEAKNQGGVI124
CL1_0213 NEENTKLFQQIYQLTGIQGVPAIAITYNGTLYAIIEGEFNV--TATPRIIQAAMSDNGVI147
GQS_02835 NEENSKLFQRIYQITGIQGVPAIAITYNGTLYAVLEGEFNV--TAVPKIIYTGMKNNGAI150
TK1208 DETNSELFGQLAQLTGITGVPAIAITYNGTIQAIFEGEFNV--SATPEIVATAMNANGVI145
A0127_08885 NETNMELFRQLNQLTGITGVPAIAIVYNDSLQAIFEGEFKV--NATPEIIATAMKENGVI145
TEU_08320 NQTNMELFNQLAQLTGITGVPAIAIAYNGTLYAVIEGEFDV--EKAPEIIATAMENNGVF147
BD01_0376 NQTNMMLFNELAQLTGITGVPAIAITYNGTLYAVIEGEFNV--TATPEIIATAIKNDGVL147
WP_010478037 NQYNIRAFNNISSLTGITGVPAIGIVYDGQLVAVIEGEFNV--SATPEIVKAAIDRNGTI128
TGAM_1636 NSHNDEVLNNISRLIGVTGVPVIGITYNGTLVAVIEGEYKV--SATPEIVETALKNNGTL129
X802_01105 NNHNINVFGNISRLTGITGVPAIGIVYNGTLVAVIEGEYNV--SATPEIIKAAIENHGML128
TON_0470 NEENQKLFGEQYKYTGIMGVPAVAITYNGTLYAIIEGEFNV--SATPEIIEAAMENNGLI125
APY94_05295 NTENQALFGEQYRYTGISGVPAIGIAYEGELRAIVEGEFNV--SAAPKIVEAAIENDGLI125
AMR53_01730 NDENNKLFSAQYKYTGIAGVPAIGIAYDGKLVAIVEGEYNV--SATPKIVQAALDNGGLI125
WP_048150379 NDENSALFGEIYRLTGISGVPAIGIAYDGKLYAIIEGEFNV--SATPEIIKEAQERNGMF125
PAP_09015 NEENGALFSEVYQLTGISGVPAIGIAYDNKLYAIIEGEFNV--SATTEIIKTAQENNGLI125
: :* .: *.. : *:.:** :: . . .
TSIB_RS09715 F-ITDKVYLIPGNNTEII---NKLESIFTNGEPSEV------174
XD43_1009 F-ITDKTYLIPENQTEII---NKLEYIFTNGEPSEV------170
OCC_10925 F-ITDKTYLIPENQTEII---NKLEYIFTNGEPSEV------170
TES1_0848 F-IADKTYLFPSNQTETI---KKLENIFMNGEPSEG------205
TERMP_RS04160 F-ITDKTYLIPLNQTETI---KKLENIFMNGEPSEG------183
PYCH_08480 V-LSSRAYLIPFNETDIV---QKLTELFLSGQASQ------165
Py04_1135 LILSSGQYILPLNNTKAESAVTNMTKWFLMGQPEP------188
PH1130 LIISQGQFLVPKNESKGLELIGNMTTWFKLNGH------170
PAB1651 LIISRGQFLIPRNETKGIEAISNLTSWFKLGEH------183
PNA2_1761 LIISSGQFIIPGNETKI---IGNLTSWFKLGEQ------166
PF0724 LVTSGKAYILKWDSQE----VQKLKEIFLSGKVNET------156
CL1_0213 LFVGGQAYILPRDKEDSAKVIDELYVLFVEHKMPDSS------184
GQS_02835 LAVGGKVYLLPRDKEDSAKVIDELYTLFVEHKMPPQESN------STSD------193
TK1208 LFIGGEWYLLARDNPRSETLIDALQTIFVEHESVDVQSVLTKLKSNSTG------194
A0127_08885 MVVQGQIYLLPRNDPRYSALIDALRTIFVEHRGVDVQSVLEKLKSNSTG------194
TEU_08320 LI-VGKVYLLPRDEPKWKLVIDALHTLFVEHEPVDVQEILDQLKANSTSAG------197
BD01_0376 LL-TGKTYLLSYKDPKAKLVINALYTIFVKHQSVDVQKIFNELKANSTSTG------197
WP_010478037 LLVGGGAYLIPHNTTGGMNLTNALYELFVRPTAT------TT164
TGAM_1636 LLLASGNYILPHNNTKAREIVEKLQYLFTKAGEPSETSTTPKK--THTTASETPADGTHT187
X802_01105 LFVGGRAYILPRNETEAMETIEKLQYLFTRAGEPSKTSTTTKK--THTTTSKATANETHI186
TON_0470 LFVAGQAYIIKNE-----TIIQKLQTIYVEHRLPEADTTETSEITTS-----TPSEDETT175
APY94_05295 LVMGGKAYIIKNE-----TIIQKLQVIYVEHRNPDETVTTTT------TP-TNN-T168
AMR53_01730 LFTGGQAYIIKNE-----TIIQELQAIYVEHRMPGEGQTTTT------TA-SETTS169
WP_048150379 LVTGGKVYLIPRNETEGAQTIAKLETLFIEHRSATGTTGTSPTTTSNTETTETTSTALPT185
PAP_09015 LFVGGKAYILKNE-----THIEMLEKLFIEHKSALNETQTSST------163
. ::. . : :
TSIB_RS09715 ------174
XD43_1009 ------170
OCC_10925 ------170
TES1_0848 ------205
TERMP_RS04160 ------183
PYCH_08480 ------165
Py04_1135 ------188
PH1130 ------170
PAB1651 ------183
PNA2_1761 ------166
PF0724 QKKAEEKSICGPASVLLLSMGAIYLRKRNYSTEVSSNV194
CL1_0213 ------184
GQS_02835 ------193
TK1208 ------194
A0127_08885 ------194
TEU_08320 ----NSTG------201
BD01_0376 ----NSTG------201
WP_010478037 TTSKGGGGICGPATVIGLSAIPILLRRRR------193
TGAM_1636 TTNGNDKRICGPALIIAIAMVPLFPRRRR------216
X802_01105 TTNGNDKRICGPALIIAIAMVPLFLGRNR------215
TON_0470 STSENNDKVCGPGIMAVLVIVPIVLLRRRR------205
APY94_05295 TTSSSDNGICGPGIVVAFAVIPLVLLKRRR------198
AMR53_01730 TTTNPGSEICGPGIAVVLAVLPLVLWKKRR------199
WP_048150379 TTGTNENKTCGPAFIGLLALLPLLMRRANH------215
PAP_09015 -QSPSKEGICGPGLIFAFALVPLLFKRRR------191
C. TON_0472 homologue
PAP_08980 ------MDELEMIRRKKMLELMKRMNGIEK--PKPKVIIEVLTSPTCPYCPIALQMAQ50
TERMP_00838 ------MDELEMIRRKKMLELMKRMGMVEVKPKGPRVVIEVITSPTCPYCPIALQMAK52
TES1_0851 ------MDELEMIRRKKMLELMKKMGMVEVKPKKPRVVIEVITSPTCPYCPIAVQMAQ52
WP_048147785 ------MDELEMIRHKKMLELMRRAGMIEEKPREPKVVIEVITSPGCPYCPIAWAMAQ52
A0127_00965 ------MDELEMIRRKKMLELMKRAGMIEAKPKGPKVIIEVITAPTCPYCPIALQMAK52
PYCH_08820 ------MDELEMIRRKKMLELMKRMGIIEEKPKRPKVTIEVITSPTCPYCPIALAMAQ52
XD43_1823 ------MIRRKKMLELMKKAGMIEEKPRGPKVVIEVITSPGCPYCPIAWAMAQ47
OCC_13031 ------MDELEMIRRKKMLELMKKAGIIEVKPKKPKVIIEVITSPTCPYCPIAWAMAQ52
TSIB_1746 MKFSWGEKMDELEMIRRKKMLELMKKAGIIEVKQEKPKVIIEVITSPGCPYCPIAWAMAQ60
PF1457 ------MIRRKKMLELMKRMGIIEERKPKKKVIIEVITAPGCPYCPIAVQMAR47
WP_010479774 ------MDELEMIRRRKMLELMKRAGMIEGN-KEPKVVIEVITSPGCPYCPIAWAMAQ51
TON_0472 ------MDELEMIRRKKMLELMKRTGMIEVKSKGPKVVIEVITAPGCPYCPIAVQMAR52
BD01_0378 ------MDELEMIRRKKMLELMKRAGMIEVKPKR-KVVIEVITAPGCPYCPIAVQMAK51
AMR53_01740 ------MDELEMIRRKKMLELMKRAGMIDVKPKR-KVVIEVITSPGCPYCPIAWAMAQ51
APY94_05305 ------MDELEMIRRKKMLELMKKAGMIEVTPKK-KVVIEVITSPGCPYCPIAWAMAQ51
CL1_0211 ------MDELEMIRRKKMLELMKRAGMIEVKPKGPKVVIEVITSPGCPYCPIAWAMAQ52
***::******:: . :: :* ***:*:* ******* **:
PAP_08980 EIGRRYQGVEVRELSVATPEGQRKAMEHNIMGVPTILINNKVEFIGVPHPGEFERKVKGY110
TERMP_00838 EIARRHPGVIVKELSVATPEGRKKAMEHNILGTPTILINNKVEFIGVPPFVEFERKVKEK112
TES1_0851 EIARKYQGIVVKELSVATPEGRKKAMEHNILGTPTILINNRVEFIGVPSFVEFERRVKEK112
WP_048147785 ELERRYEGVVAKELSVATPEGRRKAMEHNIMGTPTVLINNRVEFVGVPNFAEFERRVKRH112
A0127_00965 ELERRYEGVVAKEVSVATPEGQRKAMEHNIMGTPTILINNRVEFIGVPNFAEFESRVRAY112
PYCH_08820 EIARKYKGVVARELSIATPEGRRKAMEHNIMGTPTILINNRVEFIGVPNFAEFERKVRQY112
XD43_1823 EIARKYDGVVAKEVSVATPEGQRKAMEHNIMGTPTILINNRVEFIGVPNFAEFERRVRQY107
OCC_13031 EIARKYDGVIAKEVSVATPEGQRKAMEHNIMGTPTILINNRVEFIGVPNFAEFERRVRQY112
TSIB_1746 EIARKYDGIIAKEISVATPEGQRKAREHNIMGTPTILINNRVEFIGVPNFAEFERRVRQY120
PF1457 EIEKKYPGVIVRELSVATPEGRRKAMEHNILGTPTILINNRVEFIGVPRFIDFERRVREL107
WP_010479774 ELERKYDGVIARELSVATPEGRRKAMEHNIMGTPTILINNRVEFIGLPNFAEFERKVREL111
TON_0472 ELERKYNGVVARELSVATPEGQRKAMEHNILGTPTILINNRVEFIGVPNFGEFERKVRML112
BD01_0378 ELEGKYEGVVARELSVATPEGQRKAMEHNILGTPTILIDNRVEFIGVPNFAEFEKRVRER111
AMR53_01740 ELERKYEGVVARELSVATPEGQRKAMEHNIMGTPTILIDNRVEFIGVPNFAEFEKRVREK111
APY94_05305 ELERKYEGVIARELSVATPEGQRKAMEHNIMGTPTILIDNRVEFIGVPNFAEFERRVREK111
CL1_0211 ELERKYEGVIARELSVATPEGQRKAMEHNIMGTPTILIDNRVEFIGVPNFAEFERRVREK112
*: :: *: .:*:*:*****::** ****:*.**:**:*:***:*:* :** :*:
PAP_08980 LGI----113
TERMP_00838 LSSA---116
TES1_0851 LSSV---116
WP_048147785 LSA----115
A0127_00965 LSR----115
PYCH_08820 LSA----115
XD43_1823 LST----110
OCC_13031 LST----115
TSIB_1746 LSA----123
PF1457 LS-----109
WP_010479774 LSDNRKP118
TON_0472 LGLQ---116
BD01_0378 LGLGAPR118
AMR53_01740 LGLK---115
APY94_05305 LGLQ---115
CL1_0211 LGLQ---116
*.
D. TON_0834 homologue
PAP_08695 MIREFDGDFEKVERAKYALLWFSSPGCPPCRMIEPFMHELSEEYENVEFWEVDINKHLAL60
TON_0834 MIREFDGDFGKVERAKYALLWFSSPGCPPCRMIEPFMHELSEEYKEVEFWEVDVEKHLPL60
TES1_1390 MIREFDGDFGKVERAKYALLWFSSPGCPPCRMIEPFMHELSEEYKEVEFWEVDVGKHLWL60
Py04_0921 MIIEYEGKTNFRE--GKVVLWFSIPGCPPCRMVEGFMEELSGEFPEVKIIHVNAGEWDGL58
PF0741 MIIEYDGEIDFTK--GRVVLWFSIPGCPPCRLVERFMTELSEYFEDIQIVHINAGKWKNI58
TSIB_1703 MIHEYDGKIDFNR--GKVVLWFSIQGCPPCRLVESFMEEVSEEFSEIRFIHVGAEKWSNI58
OCC_05726 MIIEYDGKIDFTK--GKVVLWFSIPGCPPCRLVETFMEELSEEFKDIRVIHVHAEKWEDL58
A0127_10085 MIVEYDGKADFTN--GKVVLWFSIPGCPPCRLVESFMEELSDEFREIAIVHVNAEKWSEL58
TK0838 MIVEYDENVDFTK--GKAVLWFSIPGCPPCRLVEAFMKELSEEFGEIAIVHVNAEKWSGL58
WP_048147736 MIVEYDGKVDFEK--GKAVLWFSIPGCPPCRLVENFMKEVSEEFPEIKVIHVNAEEWSDL58
APY94_01785 MIVEYDGKFEFES--RKTVLWFSIPGCPPCRIVESFMEDLSGEFPEIKVVHINAEEWNDL58
AMR53_05300 MIVEYDGKINFMD--GKAVLWFSIPGCPPCRIVDSFMEELSAEFPEITVVHINAEEWNDL58
PYCH_09150 MIVEYDGKINFMD--GKAVLWFSIPGCPPCRIVDSFMEELSAEFPEITVVHINAEEWNDL58
TERMP_01376 MIVEYDGKINFMD--GKAVLWFSIPGCPPCRIVESFMEELSAEFPEITVVHINAEEWNDL58
** *:: . .:**** ******::: ** ::* : :: . .: : :
PAP_08695 IQKFDILNVPTLVYLKEGREVDRQNLIRRKEEVEERIKQLLKG-----103
TON_0834 AEKFDVMNVPTLIYLKEGNEIARQNLVRKKEEVEEKLMMLLGSDS---105
TES1_1390 AEKFDVMNVPTLIYLKEGNEITRQNLVRRREDVENILKDIIES-----103
Py04_0921 VNHFEILNIPTLVYLKDGKEMARQNLIRRKEEVLIQFEEMTRS-----101
PF0741 VDKFNILNVPTLVYLKDGREVGRQNLIRSKEEILKKLKELQE------100
TSIB_1703 VKRFEVLNVPTLVYLKDGKEVARQNLIRSKEEVLAKIEELHEC-----101
OCC_05726 VDKFDILNVPTLVYLKDGKELTRQNLIRTKEEVLLKFEELRES-----101
A0127_10085 VGKF--LNVPTLVYLKDGNEVWRQNLIREKEEVLIKLKELREL-----99
TK0838 VEGFRILNVPTLVYLKDGKEVARQNLIRGKGEVLIKFEEPREL-----101
WP_048147736 VNRFDVLNVPTLVYLKDGKEVARQNLIRNREEVLLRLEELRELQRAGL106
APY94_01785 VNRFDVLNVPTLIYLTDGKEVGRQNLIRKKEEVLIRFEELRRL-----101
AMR53_05300 VNRFDVLNVPTLIYLKDGEEVARQNLIRRK-EVLIRFEELKRL-----100
PYCH_09150 VNRFDVLNVPTLIYLKDGEEVARQNLIRRKEEVLIRFEELKRL-----101
TERMP_01376 VNRFDVLNVPTLIYLKDGEEVARQNLIRGKEEVLIRFEELKRL-----101
* :*:***:**.:*.*: ****:* : :: :
Fig. S2 Multiple sequence alignments of Trx- or Grx-like proteins. Multiple alignments were conducted using the Clustal Omega program on web server ( The locus tag or NCBI reference sequence are identical to those in Table S1 and Fig. S2.
Fig.S3SDS-PAGE analysis of the purified Trx- or Grx-like proteins, Pdo, TON_0834, and TON_0472.3 µg of proteins of Pdo and TON_0834 and 6 µg of proteins of TON_0472 were loaded onto a 15% SDS-PAGE gel. The intact proteinsand fragments of TON_0472 are marked.
Fig.S4Effect of Pdo concentration on the reduction SurR. Reduction of disulfide bonds of 4 µMSurR by 50 nM TrxR in the presence of 100 µM NADH was detected with increased concentration of Pdo All proteins (oxidized and reduced) were detected by staining with Coomassie brilliant blue as a loading control (upper panel) and mBBr-labeled proteins were detected by fluorescence illumination (lower panel) on 15% SDS-PAGE gels. M Molecular mass marker
Fig.S5Comparison of nucleotide sequences of mbh1 promotor regionsbetween T. onnurineusNA1 (TO_mbh1) and P. furiosus (PF_mbh1).The sequences of TO_mbh1 (GenBank accession no. CP000855) and PF_mbh1 (AE009950) were obtained from NCBI ( sequence alignment was conducted using the LALIGN program onweb server (
1