Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3

Atsushi Terada, Takashi Honda, Hideo Fukuhara, Kazumasa Hada, Makoto Kimura

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Ribonuclease P (RNase P) is a ribonucleoprotein complex involved in the processing of the 5′-leader sequence of precursor tRNA (pre-tRNA). Our earlier study revealed that RNase P RNA (pRNA) and five proteins (PhoPop5, PhoRpp38, PhoRpp21, PhoRpp29, and PhoRpp30) in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 reconstituted RNase P activity that exhibits enzymatic properties like those of the authentic enzyme. In present study, we investigated involvement of the individual proteins in RNase P activity. Two particles (R-3Ps), in which pRNA was mixed with three proteins, PhoPop5, PhoRpp30, and PhoRpp38 or Phopop5, PhoRpp30, and PhoRpp21 showed a detectable RNase P activity, and five reconstituted particles (R-4Ps) composed of pRNA and four proteins exhibited RNase P activity, albeit at reduced level compared to that of the reconstituted particle (R-5P) composed of pRNA and five proteins. Time-course analysis of the RNase P activities of R-4Ps indicated that the R-4Ps lacking PhoPop5, PhoRpp21, or PhoRpp30 had virtually reduced activity, while omission of PhoRpp29 or PhoRpp38 had a slight effect on the activity. The results indicate that the proteins contribute to RNase P activity in order of PhoPop5 > PhoRpp30 > PhoRpp21 ≫ PhoRpp29 > PhoRpp38. It was further found that R-4Ps showed a characteristic Mg2+ ion dependency approximately identical to that of R-5P. However, R-4Ps had optimum temperature of around at 55°C which is lower than 70°C for R-5P. Together, it is suggested that the P. horikoshii RNase P proteins are predominantly involved in optimization of the pRNA conformation, though they are individually dispensable for RNase P activity in vitro.

Original languageEnglish
Pages (from-to)293-298
Number of pages6
JournalJournal of Biochemistry
Volume140
Issue number2
DOIs
Publication statusPublished - Aug 1 2006

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Pyrococcus horikoshii
Ribonuclease P
Proteins
RNA
Nucleic Acid Conformation
Ribonucleoproteins
Archaea
RNA Precursors
Conformations

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3. / Terada, Atsushi; Honda, Takashi; Fukuhara, Hideo; Hada, Kazumasa; Kimura, Makoto.

In: Journal of Biochemistry, Vol. 140, No. 2, 01.08.2006, p. 293-298.

Research output: Contribution to journalArticle

Terada, A, Honda, T, Fukuhara, H, Hada, K & Kimura, M 2006, 'Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3', Journal of Biochemistry, vol. 140, no. 2, pp. 293-298. https://doi.org/10.1093/jb/mvj144
Terada, Atsushi ; Honda, Takashi ; Fukuhara, Hideo ; Hada, Kazumasa ; Kimura, Makoto. / Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3. In: Journal of Biochemistry. 2006 ; Vol. 140, No. 2. pp. 293-298.
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