Ribosomal protein-dependent orientation of the 16 S rRNA environment of S15.
Ribosomal protein S15 binds specifically to the central domain of 16 S ribosomal RNA (16 S rRNA) and directs the assembly of four additional proteins to this domain. The central domain of 16 S rRNA along with these five proteins form the platform of the 30 S subunit. Previously, directed hydroxyl radical probing from Fe(II)-S15 in small ribonucleoprotein complexes was used to study assembly of the central domain of 16 S rRNA. Here, this same approach was used to understand the 16 S rRNA environment of Fe(II)-S15 in 30 S subunits and to determine the ribosomal proteins that are involved in forming the mature S15-16 S rRNA environment. We have identified additional sites of Fe(II)-S15-directed cleavage in 30S subunits compared to the binary complex of Fe(II)-S15/16 S rRNA. Along with novel targets in the central domain, sites within the 5' and 3' minor domains are also cleaved. This suggests that during the course of 30S subunit assembly these elements are positioned in the vicinity of S15. Besides the previously determined role for S8, roles for S5, S6+S18, and S16 in altering the 16 S rRNA environment of S15 were established. These studies reveal that ribosomal proteins can alter the assembly of regions of the 30 S subunit from a considerable distance and influence the overall conformation of this ribonucleoprotein particle.
Jagannathan
Indu
I
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, 4258 Molecular Biology Building, Ames, IA 50011, USA.
Culver
Gloria M
GM
D001665:Binding Sites; D017665:Hydroxyl Radical; D007501:Iron / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011487:Protein Conformation; D012323:RNA Processing, Post-Transcriptional; D012336:RNA, Ribosomal, 16S / Q000737:chemistry* / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D011994:Recombinant Proteins; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C045454:ribosomal protein S11; C034212:ribosomal protein S12; C036004:ribosomal protein S15; C032643:ribosomal protein S21; C030831:ribosomal protein S5; D017665:Hydroxyl Radical; D012343:RNA, Transfer; D007501:Iron