30S ribosomal subunits can be assembled in vivo without primary binding ribosomal protein S15

• Published in: RNA (Cambridge) Vol. 12; no. 7; pp. 1229 - 1239
• Main Authors: Bubunenko, Mikhail, Korepanov, Alexey, Court, Donald L, Jagannathan, Indu, Dickinson, Daniel, Chaudhuri, Biswajoy Roy, Garber, Maria B, Culver, Gloria M
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 01.07.2006
• Subjects: Base Sequence; DNA Primers; Electrophoresis, Gel, Two-Dimensional; Escherichia coli; Escherichia coli - genetics; Models, Molecular; Molecular Sequence Data; Protein Conformation; Protein Subunits; Ribosomal Proteins - chemistry; Ribosomal Proteins - isolation & purification; Ribosomal Proteins - metabolism; RNA, Bacterial - chemistry; RNA, Bacterial - metabolism
• ISSN: 1355-8382; 1469-9001
• DOI: 10.1261/rna.2262106

Assembly of 30S ribosomal subunits from Escherichia coli has been dissected in detail using an in vitro system. Such studies have allowed characterization of the role for ribosomal protein S15 in the hierarchical assembly of 30S subunits; S15 is a primary binding protein that orchestrates the assembly of ribosomal proteins S6, S11, S18, and S21 with the central domain of 16S ribosomal RNA to form the platform of the 30S subunit. In vitro S15 is the sole primary binding protein in this cascade, performing a critical role during assembly of these four proteins. To investigate the role of S15 in vivo, the essential nature of rpsO, the gene encoding S15, was examined. Surprisingly, E. coli with an in-frame deletion of rpsO are viable, although at 37 degrees C this DeltarpsO strain has an exaggerated doubling time compared to its parental strain. In the absence of S15, the remaining four platform proteins are assembled into ribosomes in vivo, and the overall architecture of the 30S subunits formed in the DeltarpsO strain at 37 degrees C is not altered. Nonetheless, 30S subunits lacking S15 appear to be somewhat defective in subunit association in vivo and in vitro. In addition, this strain is cold sensitive, displaying a marked ribosome biogenesis defect at low temperature, suggesting that under nonideal conditions S15 is critical for assembly. The viability of this strain indicates that in vivo functional populations of 70S ribosomes must form in the absence of S15 and that 30S subunit assembly has a plasicity that has not previously been revealed or characterized.