ROLE OF THE D-ARM AND THE ANTICODON ARM IN TRANSFER-RNA RECOGNITION BY EUBACTERIAL AND EUKARYOTIC RNASE P-ENZYMES

被引:60
作者
HARDT, WD [1 ]
SCHLEGL, J [1 ]
ERDMANN, VA [1 ]
HARTMANN, RK [1 ]
机构
[1] FREE UNIV BERLIN, INST BIOCHEM, THIELALLEE 63, D-14195 BERLIN, GERMANY
关键词
D O I
10.1021/bi00211a014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Truncated precursor tRNAs lacking the D arm or anticodon arm were studied in vitro as substrates for RNase P enzymes from Escherichia coli, Thermus thermophilus (eubacteria), and HeLa. Deletion of the D arm still allowed 5'-processing by E. coli RNase P, but strongly impaired maturation by T. thermophilus and HeLa extracts. In contrast, deletion of the anticodon arm had no influence on processing by RNase P activities from all three organisms. Inhibition kinetics and gel retardation studies showed that deletion of the D arm leads to low-affinity binding to E. coli RNase P RNA (M1 RNA). However, the E. coli enzyme appears to form sufficiently strong contacts in the region of the T arm, acceptor stem, and CCA terminus to still allow productive enzyme-substrate interaction even in the absence of the structural contribution provided by the D arm. Pb2+-induced hydrolysis of a tRNA(Gly) from T. thermophilus gave identical cleavage patterns in the D arm and anticodon loop in the absence and presence of E. coli M1 RNA, whereas lead hydrolysis was strongly reduced at the CUCCAA 3'-terminus due to the presence of the enzyme.
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页码:13046 / 13053
页数:8
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