Publikationsdatum:
2014-01-07
Beschreibung:
Ribonuclease P (RNase P) is a ubiquitous trans-acting ribozyme that processes the 5' leader sequence of precursor tRNA (pre-tRNA). The secondary structure of RNase P RNA ( Pho pRNA) in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 has several peripheral stem-loops. In order to investigate their functional role, we prepared six mutants, P1, P3, P8, P9, P12 and P15, in which the stem-loops including helices P1, P3, P8, P9, P12/12.1/12.2 and P15/16 in Pho pRNA were individually deleted, respectively, and characterized them with respect to pre-tRNA cleavage activity in the presence of five proteins and also to the ability to form a complex with the proteins. The reconstituted particles containing P3, P8 or P9 retained considerable levels of activity (35–65%), while those containing P1, P12 or P15 had markedly reduced activity (13%). It was further found that the reconstituted particles comprising P3 or P15 lacked Pho Pop5 and Pho Rpp30, whereas those containing P1, P8, P9 or P12 bound to all five proteins. Since it is known that Pho Pop5 functions in a complex with Pho Rpp30, the present result suggests that the peripheral stem-loops containing P3 or P15/16 are involved in the structural formation of a catalytic site by interacting with the protein complex Pho Pop5- Pho Rpp30.
Print ISSN:
0021-924X
Digitale ISSN:
1756-2651
Thema:
Biologie
,
Chemie und Pharmazie
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