ACA-specific RNA sequence recognition is acquired via the loop 2 region of MazF mRNA interferase.
ACA-specific RNA sequence recognition is acquired via the loop 2 region of MazF mRNA interferase.
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DOI：
10.1002/prot.24246
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发表时间：
2013-05
影响因子：
2.9
通讯作者：
Inouye, Masayori
中科院分区：
文献类型：
--
作者：
Park, Jung-Ho;Yoshizumi, Satoshi;Yamaguchi, Yoshihiro;Wu, Kuen-Phon;Inouye, Masayori
MazF是一种mRNA干扰酶，它在特定的RNA序列上切割mRNA。来自大肠杆菌的MazF （MazF-ec）在A^CA切割RNA。迄今为止，已经从细菌到古细菌中鉴定出大量能够切割特定3到7个碱基序列的MazF同源物。MazF-ec形成二聚体，其中两个亚基之间的界面已知是RNA底物结合位点。在这里，我们研究了两个环在MazF-ec中的作用，这两个环与MazF-ec二聚体的界面密切相关。我们检测了将MazF-ec的环区与其他MazF同源物(如来自黄粘球菌（MazF-mx）和来自结核分枝杆菌（MazF-mt3）的MazF交换是否会影响RNA切割的特异性。我们发现，将MazF-ec的环2与MazF-mx或MazF-mt3的环2区域交换，除了原始的切割位点a ^CA外，还在（a /U）（a /U）AA^C处产生了一个新的切割序列，而交换环1并没有改变切割的特异性。有趣的是，环2与8或12个连续的Gly残基交换也会在（a /U）（a /U）AA^C上产生一个新的RNA切割位点。本研究提出了一种扩大mRNA干扰酶的RNA切割库的方法，这对特定基因表达的调控和生物技术应用的潜在用途至关重要。
MazF is an mRNA interferase that cleaves mRNAs at a specific RNA sequence. MazF from E. coli (MazF-ec) cleaves RNA at A^CA. To date, a large number of MazF homologues that cleave RNA at specific three- to seven-base sequences have been identified from bacteria to archaea. MazF-ec forms a dimer, in which the interface between the two subunits is known to be the RNA substrate-binding site. Here, we investigated the role of the two loops in MazF-ec, which are closely associated with the interface of the MazF-ec dimer. We examined whether exchanging the loop regions of MazF-ec with those from other MazF homologues, such as MazF from Myxococcus xanthus (MazF-mx) and MazF from Mycobacterium tuberculosis (MazF-mt3), affects RNA cleavage specificity. We found that exchanging loop 2 of MazF-ec with loop 2 regions from either MazF-mx or MazF-mt3 created a new cleavage sequence at (A/U)(A/U)AA^C in addition to the original cleavage site, A^CA, while exchanging loop 1 did not alter cleavage specificity. Intriguingly, exchange of loop 2 with 8 or 12 consecutive Gly residues also resulted in a new RNA cleavage site at (A/U)(A/U)AA^C. The present study suggests a method for expanding the RNA cleavage repertoire of mRNA interferases, which is crucial for potential use in the regulation of specific gene expression and for biotechnological applications.
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影响因子：
5.6
作者：
Kamphuis, MB;Bonvin, AMJJ;Boelens, R
通讯作者：
Boelens, R
影响因子：
16.6
作者：
Yamaguchi, Yoshihiro;Nariya, Hirofumi;Inouye, Masayori
通讯作者：
Inouye, Masayori
DOI：
10.1093/protein/8.2.127
发表时间：
1995-02-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
WALLACE, AC;LASKOWSKI, RA;THORNTON, JM
通讯作者：
THORNTON, JM
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
4.8
作者：
Zhang, JJ;Zhang, YL;Inouye, M
通讯作者：
Inouye, M