Rubra_00390 Rubra_00390   Rubra_00410 Rubra_00410

Rubra_00400 : CDS information

close this sectionLocation

Organism
StrainNRRL 3061 (=NBRC 14000)
Entry nameRubradirin
Contig
Start / Stop / Direction77,653 / 81,606 / + [in whole cluster]
77,653 / 81,606 / + [in contig]
Location77653..81606 [in whole cluster]
77653..81606 [in contig]
TypeCDS
Length3,954 bp (1,317 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category1.4 Other
Productaminocoumarin--carboxylic acid ligase/L-tyrosine--PCP ligase/peptidyl carrier protein
amide synthetase/L-tyrosyl-PCP synthetase/peptidyl carrier protein
Product (GenBank)putative CoA ligase
Gene
Gene (GenBank)rubC1
EC number
Keyword
  • aminocoumarin moiety
Note
Note (GenBank)
Reference
ACC
PmId
[18080113] Biosynthesis of rubradirin as an ansamycin antibiotic from Streptomyces achromogenes var. rubradiris NRRL3061. (Arch Microbiol. , 2008)
[21448870] Adenylate-forming enzymes of rubradirin biosynthesis: RubC1 is a bifunctional enzyme with aminocoumarin acyl ligase and tyrosine-activating domains. (Chembiochem. , 2011)
[23225745] A domain of RubC1 of rubradirin biosynthesis can functionally replace MbtH-like proteins in tyrosine adenylation. (Chembiochem. , 2013)
comment
[PMID: 18080113](2008)
Rubradirin生合成clusterの報告。
rubC1-C5はAMC and DHDP biosynthesisに関連する。

Table1>配列比較からの推定機能
RubC1(1317aa): CoA ligase

rubC1-C4はaminocoumarin生合成に関連するものに似ている。RubC1は、アミド結合の形成で機能するNovLと、tyrosineのadenylationで機能するNovHにAA配列類似性を示す。NovLとNovHは分離したタンパクとして同定されているが、rubC1ではNovL and NovH domainsの両方を持った1つの遺伝子として検出された。

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[PMID:21448870](2011)
aminocoumarin生合成、結合に関連するであろうOrf4, RubF6, RubC1を発現、精製して調査している。
RubC1は、aminocoumarin acyl ligase と tyrosine-activating domains (tyrosine-adenylation and peptidyl-carrier domains)とのbifunctional enzymeである。

ruburadurun生合成において、Orf4と、RubC1のN末domainの両方がaminocoumarin acyl ligasesとして働くことができるかもしれない。
RubC1, C2, C3の並びはnovobiocin生合成clusterでのsingle operon NovHIJKLに似ていることから、RubC1のほうが効率的でありうる。
Related Reference
ACC
Q9L9F6
PmId
[10801869] Cloning, overexpression, and purification of novobiocic acid synthetase from Streptomyces spheroides NCIMB 11891. (J Biol Chem. , 2000)
comment
Streptomyces sphaeroides_novL
NovL

blast search
Rubra_00110 2e-70 260 36.01 <ORF4
Rubra_00400 4e-67 249 37.5 <this ORF, query N末側半分にhit
Rubra_00310 7e-66 245 35.26 <RubF6

---
Novobiocic acid synthetaseをコードするnovLをE.coliで発現、精製、活性測定。
3-dimethylallyl-4-hydroxybenzoic acid (ring A of novobiocin) と
3-amino-4,7-dihydroxy-8-methyl coumarin (ring B of novobiocin)
の間のアミド結合の形成をATP-dependent reactionで触媒した。
また、ring A and ATPからのacyl adenylateの形成がATP-PP(i) exchange assayによって実証された。
すなわちNovLは、acyl adenylateのようなring Aの活性化と、引き続くacyl基のring Bのamino基への移動の両方を触媒した。
ACC
Q9L9G0
PmId
[11325587] Coumarin formation in novobiocin biosynthesis: beta-hydroxylation of the aminoacyl enzyme tyrosyl-S-NovH by a cytochrome P450 NovI. (Chem Biol. , 2001)
comment
12th(Q9L9G0) 48%, 1e-133, query C末側半分にhit
Streptomyces sphaeroides_novH
NovH

E.coliでNovHとNovIを過剰発現、精製、特徴づけ。

NovHはadenylation (A) and a peptidyl carrier protein (PCP) domainを持ち、NRPSのような構成である(Condensation domainはない)。

A domainの基質がL-Tyrであること(ATP-[32P]PPi assays)、phosphopantetheinyltransferase (PPTase)_Sfpでの処理とATPがNovHのself aminoacylationに必要であることを実験で確認している。

そこから提唱されるNovHの機能は以下の通り。
NovHはapo proteinであり、PPTaseとCoAで反応することによりholo proteinとなる。
A domainがL-tyrosyl-AMPとしてL-Tyrを活性化し、そのL-tyrosyl基をHS-pant-PCP domainへ移してL-tyrosyl-S-NovHができる。

close this sectionPKS/NRPS Module

B1 tyrosine
A757..1148
PCP1235..1301

close this sectionSequence

selected fasta
>aminocoumarin--carboxylic acid ligase/L-tyrosine--PCP ligase/peptidyl carrier protein [putative CoA ligase]
MRGHDHYIRRILGRLDAHPAGVPFVLGNDPFPAERLARAIRTTAGWMRAAGVGPGTSVAV
LTSPNTPSTLVYRYAVNLLGATAVHIRGVNAADPQDELGPRVQAEILAGLRPSVLAVDQD
NLDRARELRAAAGSAFALAAPGPYGPDVLDMSRPLESPFDDATAERAEIAVVTFTSGSSG
RPKGVCWPFDVKDDMASAAAGAQPAVCLITGTLTHSSGFSADDAIIAGGSVVLHHGFDAE
AVLRAVERHRVTRLVLASAQVYALTEHHAFDDYDRSSLREVFYTGSPAAPERLAAAAKSL
GPVLFQVYGTSETGMISLLTPQDHLDPDLRRTVGRPPANVRITIRDPRDHDRLLPPGVSG
EICSSGRWAMSHYWNDPEQTARTVRDGWVRTGDIGRLDESGYLTLEGRLDGVLKGHGVRI
HPEAIERVLLEHADVAQAAVFGIEDEDFLALVHAVVTPAPGCTPRPADLGAYVADALGDR
HAPVDIEVRSELPLLGSAKPDRNLLREQALGARFWHASLGGVDGDSLLRDRLARPGDDTL
GAPALQRRIPDFPSAKNTLLGVWAVLLHLLGAGSPVVFGAGPRPLLVAVAPGEEIGAVLH
RVEEAVAASAGHARLGPRGRAELDAAADTLFDTALTFTRVGDPPPGPARPLTAELRVGHD
AGDLVLTLVTDPRRIGADCAGRLLEMVLRLLGAFREAPGQPVASLDLLDPATRRQVLQEW
NSSRQEQPAATLGRLWQDAADAHAGRPAVEEAGVTTSYAELDRRAGRLAAAIAQAGAAPG
RMVALLLPRSLDLITALLATVRTGAAFVPIDPGYPPERITAMLEDCDPVAVCTTASADMP
STRPWPRVHLDGPEAGEPAGGFPRVAPDVADPAYVIYTSGTTGRPKGVVVTHAGLANLAA
AKREGLGLDSTARVLQFASPSFDAFVAELLGAFTSGATVVVPPQGPLAGEPLTAVLTERR
ITHAILPPVALSSMDGAAGALPGLRGLISAGEECPAELAARWSQGRRMVNAYGPTEVTVC
ATQSGPLTADGRPPIGRPVAGARAYVLGPGLQPVPPGFRGELYIAGPGVARGYLNQPALT
AVRFVADPFGPPGSRMYRTGDIASWRADGNLDFHGRADDQIKLRGFRIEPREVAAVLEEL
PAVARAVAGVREDRGGRARLIAWVVPADGEAPTPERLREHAARRLPEHMTPTGYVLVSAL
PMTPNGKLDLAALPAVAERDSPPPAAARPALRTPAERALAAIVGELLGATDVGADSDFFA
LGGDSMLAISLIRRAREAGLALSPKEIVGNPTIGALAAVATSLPSPPAQTPHVEGRR
selected fasta
>aminocoumarin--carboxylic acid ligase/L-tyrosine--PCP ligase/peptidyl carrier protein [putative CoA ligase]
TTGCGGGGGCATGATCACTACATCCGCCGGATCCTCGGCCGGCTCGACGCCCATCCTGCC
GGCGTTCCCTTCGTGCTCGGAAACGACCCCTTCCCGGCCGAGCGTCTCGCCCGTGCGATC
CGCACGACCGCCGGGTGGATGCGCGCGGCCGGTGTCGGCCCGGGCACCTCGGTCGCCGTG
CTGACCAGTCCCAACACACCGTCCACCCTGGTGTACCGGTACGCGGTGAACCTCCTGGGC
GCGACGGCGGTCCACATTCGCGGGGTGAACGCCGCGGACCCTCAGGACGAGTTGGGTCCT
CGCGTCCAGGCGGAGATTCTCGCCGGGCTGCGGCCGAGCGTGCTGGCGGTGGACCAGGAC
AACCTCGACCGTGCGCGCGAGCTGCGTGCCGCGGCCGGATCGGCGTTCGCCCTCGCCGCC
CCGGGTCCGTACGGACCGGATGTTCTGGACATGAGCCGCCCCTTGGAGTCGCCGTTCGAC
GACGCGACGGCCGAGCGGGCGGAGATCGCCGTGGTGACGTTCACCAGCGGATCGTCGGGA
CGGCCCAAGGGCGTCTGCTGGCCGTTCGACGTGAAGGACGACATGGCCTCGGCGGCCGCC
GGGGCCCAGCCGGCGGTCTGTCTGATCACCGGGACGCTGACCCACTCCAGCGGTTTCTCG
GCCGATGACGCGATCATCGCCGGGGGCAGTGTCGTGCTGCACCACGGGTTCGACGCCGAG
GCGGTGCTCCGAGCCGTCGAACGGCACCGCGTCACCCGGCTCGTCCTCGCCTCTGCGCAG
GTCTACGCCCTCACCGAGCATCACGCGTTCGACGACTACGACCGCTCCAGCCTGCGCGAG
GTGTTCTACACCGGGAGCCCGGCCGCGCCGGAGCGGCTGGCGGCCGCCGCCAAATCGCTC
GGCCCGGTGCTGTTCCAGGTCTACGGCACGAGCGAGACCGGCATGATCAGCCTGCTCACC
CCACAGGATCATCTGGACCCCGACCTGCGGCGCACGGTCGGCCGGCCCCCGGCGAACGTC
CGGATCACCATCCGTGATCCGCGAGACCACGACCGGCTCCTCCCGCCGGGCGTCTCCGGC
GAGATCTGCTCGTCCGGGCGCTGGGCGATGAGCCACTACTGGAACGACCCGGAGCAGACC
GCGCGGACGGTCCGCGACGGGTGGGTGCGCACCGGCGACATCGGACGGCTCGACGAGTCG
GGATACCTGACCCTCGAGGGCCGCCTCGACGGGGTCCTGAAGGGTCACGGTGTCCGCATC
CACCCGGAGGCGATCGAGCGCGTGCTCCTCGAACACGCCGACGTGGCCCAGGCCGCGGTG
TTCGGGATCGAGGACGAGGATTTCCTGGCGCTGGTCCACGCGGTGGTGACGCCGGCGCCC
GGCTGCACGCCGCGGCCGGCCGACCTGGGGGCGTACGTGGCGGACGCGCTCGGTGACCGC
CATGCTCCGGTGGACATCGAGGTCAGGTCCGAACTGCCGCTGCTGGGATCGGCGAAGCCG
GACCGGAACCTGCTGCGCGAGCAGGCGCTGGGCGCGCGGTTCTGGCACGCGTCGCTAGGG
GGGGTCGACGGTGATTCCCTGCTCCGGGACAGGCTCGCCCGGCCGGGCGACGACACGTTG
GGGGCACCCGCACTCCAGCGGCGGATCCCGGATTTCCCGTCCGCAAAGAACACGCTGCTC
GGAGTCTGGGCGGTCCTGCTGCACCTGCTGGGCGCCGGCTCGCCGGTCGTCTTCGGGGCC
GGGCCCCGGCCGCTGCTGGTGGCCGTCGCGCCGGGCGAGGAGATCGGCGCCGTCCTCCAC
CGGGTCGAGGAGGCCGTGGCCGCCTCGGCCGGGCACGCCCGGCTGGGACCGCGGGGGCGG
GCCGAACTCGACGCCGCGGCAGACACGTTGTTCGACACCGCGCTCACCTTCACCCGTGTC
GGGGACCCGCCGCCCGGCCCGGCCCGGCCGCTCACGGCCGAGCTCAGGGTGGGCCACGAC
GCGGGCGATCTCGTCCTGACCCTGGTCACCGACCCGCGGCGGATCGGCGCGGACTGCGCC
GGACGCCTCCTCGAGATGGTCCTGCGCCTGCTCGGGGCATTTCGCGAGGCGCCCGGCCAG
CCGGTCGCCTCGCTGGACCTCCTCGACCCGGCCACCCGCCGCCAGGTGCTGCAGGAATGG
AACTCCTCCCGGCAGGAGCAGCCCGCCGCGACCCTGGGCCGGCTGTGGCAGGACGCTGCG
GACGCACACGCCGGACGGCCGGCCGTGGAAGAGGCCGGCGTCACCACCTCGTACGCCGAA
CTCGACCGCCGTGCCGGCCGGCTCGCGGCGGCCATCGCGCAGGCGGGTGCGGCGCCGGGC
CGGATGGTGGCCCTCCTGCTGCCGCGGTCCCTGGACCTGATCACCGCGCTCCTGGCGACC
GTCCGGACCGGCGCGGCGTTCGTCCCCATCGACCCGGGATACCCGCCAGAGCGGATCACG
GCGATGCTGGAGGACTGCGACCCGGTGGCGGTCTGCACGACGGCGTCGGCCGACATGCCC
TCGACCCGTCCCTGGCCGCGCGTCCACCTGGACGGGCCGGAGGCCGGCGAGCCGGCGGGC
GGGTTTCCCCGGGTCGCGCCCGACGTGGCCGATCCGGCGTACGTCATCTACACCTCCGGC
ACGACCGGCCGGCCGAAGGGCGTCGTGGTCACCCACGCCGGGCTGGCGAACCTGGCCGCG
GCCAAACGCGAGGGCCTGGGACTGGACAGCACCGCCCGGGTCCTGCAGTTCGCCTCACCG
AGCTTCGACGCGTTCGTCGCCGAGCTGTTGGGCGCGTTCACCTCGGGGGCGACGGTGGTC
GTGCCGCCGCAGGGCCCCCTCGCCGGAGAACCGCTGACGGCGGTTCTGACCGAGCGCCGG
ATCACCCACGCCATCCTGCCGCCGGTGGCACTGTCCAGCATGGACGGCGCCGCCGGCGCA
CTGCCCGGGCTCCGCGGCCTGATCAGCGCGGGCGAGGAGTGCCCGGCCGAGCTGGCCGCG
CGCTGGTCGCAGGGCCGCCGGATGGTCAACGCCTACGGGCCCACCGAGGTCACCGTCTGC
GCCACTCAGAGCGGGCCGCTCACGGCAGACGGCCGGCCTCCGATCGGACGGCCCGTCGCC
GGTGCCCGCGCCTACGTGCTCGGCCCCGGGCTGCAGCCGGTGCCACCCGGATTCCGCGGC
GAGCTCTACATCGCGGGCCCCGGCGTCGCCCGGGGTTACCTCAACCAGCCCGCCCTGACA
GCCGTACGCTTCGTGGCGGACCCGTTCGGCCCGCCCGGCAGCCGGATGTACCGCACCGGC
GACATCGCCTCCTGGCGCGCCGACGGCAACCTGGACTTCCACGGCCGGGCCGACGACCAG
ATCAAACTGCGGGGCTTTCGGATCGAGCCGCGCGAGGTGGCGGCGGTGCTGGAGGAACTG
CCCGCGGTCGCCCGTGCGGTGGCCGGCGTACGCGAGGACCGTGGCGGCCGGGCCCGGCTG
ATCGCCTGGGTCGTGCCCGCCGACGGGGAGGCACCCACCCCCGAGCGGCTCCGGGAGCAC
GCGGCACGCCGGCTGCCGGAGCACATGACGCCGACGGGGTACGTCCTCGTCAGTGCCCTG
CCCATGACGCCGAACGGCAAGCTGGACCTGGCCGCGCTGCCCGCGGTCGCCGAGCGGGAC
AGTCCCCCGCCCGCGGCGGCGCGGCCGGCCCTGCGCACGCCGGCGGAACGCGCGCTGGCC
GCCATCGTCGGGGAGCTGCTGGGCGCCACGGACGTCGGCGCCGACAGCGACTTCTTCGCG
CTCGGCGGCGACAGCATGCTGGCCATCTCGCTGATCCGGCGCGCCCGCGAAGCGGGTCTG
GCCCTCTCCCCCAAGGAGATCGTGGGAAACCCCACGATCGGTGCCCTCGCCGCGGTGGCC
ACCTCGCTGCCGTCCCCGCCCGCCCAGACGCCTCACGTGGAAGGCCGCCGATGA
[1] A757..1148
[1] tyrosine923..1020
[1] PCP1235..1301
[1] A2269..3444
[1] tyrosine2767..3060
[1] PCP3703..3903

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR000873 AMP-dependent synthetase/ligase (Domain)
[36-440] 7.4e-63 PF00501 [757-1147] 1.60000000000002e-106 PF00501
PF00501   AMP-binding
IPR009081 Acyl carrier protein-like (Domain)
[1228-1302] 1.39999892049878e-15 SSF47336
SSF47336   ACP_like
[1229-1302] 5.80000000000004e-21 G3DSA:1.10.1200.10
G3DSA:1.10.1200.10   ACP_like
[1235-1301] PS50075
PS50075   ACP_DOMAIN
[1238-1298] 7.7e-13 PF00550
PF00550   PP-binding
IPR010071 Amino acid adenylation domain (Domain)
[757-1148] 2.20000000000002e-123 TIGR01733
TIGR01733   AA-adenyl-dom
IPR020845 AMP-binding, conserved site (Conserved_site)
[172-183] PS00455 [875-886] PS00455
PS00455   AMP_BINDING
SignalP
[1-27] 0.136 Signal
Eukaryota   
TMHMM No significant hit
Rubra_00390 Rubra_00390   Rubra_00410 Rubra_00410
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