Ansam_00110 Ansam_00110   Ansam_00130 Ansam_00130

Ansam_00120 : CDS information

close this sectionLocation

Organism
StrainATCC 31565
Entry nameAnsamitocin
Contig
Start / Stop / Direction27,437 / 26,553 / - [in whole cluster]
55,310 / 56,194 / + [in contig]
Locationcomplement(26553..27437) [in whole cluster]
55310..56194 [in contig]
TypeCDS
Length885 bp (294 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category3.2 modification methylation
ProductSAM-dependent N-methyltransferase
Product (GenBank)methyltransferase
Gene
Gene (GenBank)asm10
EC number2.1.1.-
Keyword
Note
Note (GenBank)
Reference
ACC
PmId
[12060743] The biosynthetic gene cluster of the maytansinoid antitumor agent ansamitocin from Actinosynnema pretiosum. (Proc Natl Acad Sci U S A. , 2002)
[14624546] The post-polyketide synthase modification steps in the biosynthesis of the antitumor agent ansamitocin by Actinosynnema pretiosum. (J Am Chem Soc. , 2003)
[21681880] N-methylation of the amide bond by methyltransferase asm10 in ansamitocin biosynthesis. (Chembiochem. , 2011)
comment
[PMID:12060743](2002)
Actinosynnema pretiosum ssp. auranticum ATCC 31565のcosmid libraryから、ansamitocin生合成に必要な2つのgene clustersをクローニング。

<cluster I>
Asm10(294aa) : Methyltransferase

配列解析のみ。
asm11 and asm12とpolycistronicに転写されるようだ。

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[PMID:14624546](2003)
Actinosynnema pretiosumのansamitocin生合成gene clusterにある6genesの機能を、遺伝子不活化とmutantの化学分析によって調査。

それらはそれぞれ
halogenase (asm12),
carbamoyltransferase (asm21),
20-O-methyltransferase (asm7),
3-O-acyltransferase (asm19),
epoxidase (asm11),
N-methyltransferase (asm10)
をコードし、ansamitocin形成において6つのPKS後修飾ステップを担う。

Asm7 deletion mutantの主要産物は20-O,N-didemethyl-Ansamitocin P-3、
Asm10 deletion mutantの主要産物はN-demethyl-Ansamitocin P-3であったことから、これらmethyltransferasesの割り当てが確定している。

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同じグループがAsm10について詳細に調べた続報。

[PMID:21681880](2011)
mutant作成、相補確認から、Asm10はansamitocin生合成に関与する。
in vitroでの酵素活性測定では、Asm10がSAM-dependent N-methylationを担うことと、その基質柔軟性が示された。
site-directed mutagenesisを通してAsp154 and Leu155が触媒に必須であることが確認された。

close this sectionSequence

selected fasta
>SAM-dependent N-methyltransferase [methyltransferase]
MSLPADSPPPGPVEMPDESWTSLGNAGARAQESARADRLFDDPLARAFLDAAGTGNPLLP
DRGGGDPGLLGQITDVIVVKTVFFDAVLARAAAAGVRQVVLLAAGLDARAFRLPWPEGVV
VFEVDLPDVLGFKERVVREVGAEPSCDRRVVAADLRSDWVAALVAAGLDREAPVAWLAEG
ALGLLDEAGCEELMAAVLGASAAGSRFALDHTHDGWKAGEALGGYLEGTGVCLADLVKGG
PREPGGAWLARHGWRVAEYDVVAEAARHGRPAPTLFRVPERRANATILFEAELG
selected fasta
>SAM-dependent N-methyltransferase [methyltransferase]
ATGAGCCTCCCAGCGGACTCACCACCGCCGGGGCCGGTGGAGATGCCCGACGAGAGCTGG
ACGTCGCTGGGCAACGCGGGGGCGCGGGCGCAGGAGTCCGCGCGGGCCGACCGGTTGTTC
GACGACCCGCTGGCGCGGGCGTTCCTGGACGCGGCGGGGACGGGGAACCCGCTGCTGCCC
GACCGGGGTGGCGGTGATCCGGGGCTGCTCGGGCAGATCACGGACGTCATCGTGGTCAAG
ACGGTGTTCTTCGACGCGGTGCTGGCGCGGGCGGCGGCGGCCGGGGTGCGGCAGGTGGTG
CTGCTGGCCGCCGGGTTGGACGCCCGCGCGTTCCGGCTGCCCTGGCCCGAGGGGGTGGTG
GTGTTCGAGGTGGACCTGCCGGACGTGCTGGGGTTCAAGGAGCGCGTGGTGCGGGAGGTC
GGGGCGGAGCCCTCGTGCGACCGGCGGGTCGTCGCGGCGGACCTGCGCTCGGACTGGGTG
GCCGCGCTGGTCGCCGCCGGGCTGGACCGGGAGGCCCCGGTGGCCTGGCTGGCGGAGGGC
GCGCTGGGGCTGCTGGACGAGGCCGGGTGCGAGGAGCTGATGGCGGCGGTGCTCGGGGCC
TCCGCGGCGGGGAGCCGGTTCGCGCTCGACCACACCCACGACGGGTGGAAGGCCGGGGAG
GCGCTGGGCGGGTACCTGGAGGGGACCGGGGTCTGCTTGGCCGACCTGGTGAAGGGGGGT
CCGCGGGAGCCGGGCGGGGCGTGGTTGGCGCGGCACGGGTGGCGGGTGGCGGAGTACGAC
GTCGTGGCGGAGGCGGCACGGCACGGGCGGCCCGCGCCGACCCTGTTCCGGGTGCCGGAG
CGGCGGGCGAACGCGACGATCCTGTTCGAGGCCGAGCTGGGGTAG

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR007213 Leucine carboxyl methyltransferase (Family)
[23-197] 6.40000000000004e-48 PF04072
PF04072   LCM
IPR011610 S-adenosyl-L-methionine-dependent methyltransferase, putative (Family)
[20-271] 2.09999999999996e-82 TIGR00027
TIGR00027   Mthyl_TIGR00027
SignalP
[1-36] 0.103 Signal
Bacteria, Gram-negative   
TMHMM No significant hit
Ansam_00110 Ansam_00110   Ansam_00130 Ansam_00130
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