Structure-function analysis of yeast RNA debranching enzyme (Dbr1), a manganese-dependent phosphodiesterase.
Structure-function analysis of yeast RNA debranching enzyme (Dbr1), a manganese-dependent phosphodiesterase.
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酵母RNA脱支酶（DBR1）的结构 - 功能分析，锰依赖性磷酸二酯酶。
DOI：
10.1093/nar/gki934
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发表时间：
2005
影响因子：
14.9
通讯作者：
Schwer, B
中科院分区：
文献类型：
--
作者：
Khalid, MF;Damha, MJ;Shuman, S;Schwer, B
Saccharomyces cerevisiae Dbr1 is a 405-amino acid RNA debranching enzyme that cleaves the 2′-5′ phosphodiester bonds of the lariat introns formed during pre-mRNA splicing. Debranching appears to be a rate-limiting step for the turnover of intronic RNA, insofar as the steady-state levels of lariat introns are greatly increased in a Δdbr1 strain. To gain insight to the requirements for yeast Dbr1 function, we performed a mutational analysis of 28 amino acids that are conserved in Dbr1 homologs from other organisms. We identified 13 residues (His13, Asp40, Arg45, Asp49, Tyr68, Tyr69, Asn85, His86, Glu87, His179, Asp180, His231 and His233) at which alanine substitutions resulted in lariat intron accumulation in vivo. Conservative replacements at these positions were introduced to illuminate structure–activity relationships. Residues important for Dbr1 function include putative counterparts of the amino acids that comprise the active site of the metallophosphoesterase superfamily, exemplified by the DNA phosphodiesterase Mre11. Using natural lariat RNAs and synthetic branched RNAs as substrates, we found that mutation of Asp40, Asn85, His86, His179, His231 or His233 to alanine abolishes or greatly diminishes debranching activity in vitro. Dbr1 sediments as a monomer and requires manganese as the metal cofactor for debranching.
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影响因子：
4.5
作者：
Coombes, CE;Boeke, JD
通讯作者：
Boeke, JD
影响因子：
16.8
作者：
Coppins, RL;Silverman, SK
通讯作者：
Silverman, SK
DOI：
10.1006/bbrc.1999.2048
发表时间：
2000-02-05
影响因子：
3.1
作者：
Karst, SM;Rütz, NL;Menees, TM
通讯作者：
Menees, TM
影响因子：
56.9
作者：
Cheng, Z;Menees, TM
通讯作者：
Menees, TM
影响因子：
64.8
作者：
GOLDBERG, J;HUANG, HB;KURIYAN, J
通讯作者：
KURIYAN, J