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
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.