Independent in vitro assembly of all three major morphological parts of the 30S ribosomal subunit of Thermus thermophilus

Citation
Sc. Agalarov et al., Independent in vitro assembly of all three major morphological parts of the 30S ribosomal subunit of Thermus thermophilus, EUR J BIOCH, 266(2), 1999, pp. 533-537
Citations number
22
Categorie Soggetti
Biochemistry & Biophysics
Journal title
EUROPEAN JOURNAL OF BIOCHEMISTRY
ISSN journal
00142956 → ACNP
Volume
266
Issue
2
Year of publication
1999
Pages
533 - 537
Database
ISI
SICI code
0014-2956(199912)266:2<533:IIVAOA>2.0.ZU;2-J
Abstract
Fragments of the 16S rRNA of Thermus thermophilus representing the 3' domai n (nucleotides 890-1515) and the 5' domain (nucleotides 1-539) have been pr epared by transcription in vitro. Incubation of these fragments with total 30S ribosomal proteins of T. thermophilus resulted in formation of specific RNPs. The particle assembled on the 3' RNA domain contained seven proteins corresponding to Escherichia coli ribosomal proteins S3, S7, S9, S10, S13, S14, and S19. All of them have previously been shown to interact with the 3' domain of the 16S RNA and to be localized in the head of the 30S ribosom al subunit. The;particle formed on the 5' RNA domain contained five ribosom al proteins corresponding to E. coli proteins S4, S12, S17, S16, and S20. T hese proteins are known to be localized in the main part of the body of the 30S subunit. Both types of particle were compact and had sedimentation coe fficients of 15.5 S and 13 S, respectively. Together with our recent demons tration of the reconstitution of the RNA particle representing the platform of the T. thermophilus 30S ribosomal subunit [Agalarov, S.C., Zheleznyakov a, E.N., Selivanova, O.M., Zheleznaya; L.A., Matvienko, N.I., Vasiliev, V.D . & Spirin, A.S. (1998) Proc. Natl Acad. Sci. USA 95, 999-1003], these-expe riments establish that all three main structural lobes of the small ribosom al subunit can be reconstituted independently of each other and prepared in the individual state.