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EcoSal Plus

Domain 4:

Synthesis and Processing of Macromolecules

Assembly of the 30S Ribosomal Subunit

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  • Authors: Gloria M. Culver1, and Narayanaswamy Kirthi2
  • Editor: Susan T. Lovett3
  • VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA; 2: Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA; 3: Brandeis University, Waltham, MA
  • Received 22 August 2007 Accepted 07 November 2007 Published 31 January 2008
  • Address correspondence to Gloria M. Culver gculver@mail.rochester.edu.
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  • Abstract:

    Protein synthesis involves nearly a third of the total molecules in a typical bacterial cell. Within the cell, protein synthesis is performed by the ribosomes, and research over several decades has investigated ribosomal formation, structure, and function. This review provides an overview of the current understanding of the assembly of the 30S ribosomal subunit. The 30S subunit contains one rRNA molecule (16S) and 21 ribosomal proteins (r-proteins; S1 to S21). The formation of functional subunits can occur as a self-assembly process in vitro; i.e., all the information required for the formation of active ribosomes resides in the primary sequences of the r-proteins and rRNAs. In vitro reconstitution of functional 30S subunits is carried out by using a mixture of TP30, individually purified natural or recombinant r-proteins, and natural 16S rRNA. Chemical probing and primer extension analysis have been used extensively to monitor changes in the reactivities of nucleotides in 16S rRNA during the in vitro reconstitution of 30S subunits. The potential roles for r-proteins in 30S subunit assembly were determined by omitting single proteins in reconstitution experiments. The RNPs resulting from single protein omissions were examined in terms of their composition and function to determine the roles of the absent proteins. Recent developments in understanding the structure of the 30S subunit have led to speculation about roles for some of the r-proteins in assembly. The crystal structures of the 30S subunit ( 1 , 2 ) and the 70S ribosome ( 3 ) reveal details of the r-protein and rRNA interactions.

  • Citation: Culver G, Kirthi N. 2008. Assembly of the 30S Ribosomal Subunit, EcoSal Plus 2008; doi:10.1128/ecosalplus.2.5.3

Key Concept Ranking

16s rRNA Sequencing
0.3876929
RNA Helicase
0.3387978
Thermus thermophilus
0.31044513
0.3876929

References

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/content/journal/ecosalplus/10.1128/ecosalplus.2.5.3
2008-01-31
2017-04-26

Abstract:

Protein synthesis involves nearly a third of the total molecules in a typical bacterial cell. Within the cell, protein synthesis is performed by the ribosomes, and research over several decades has investigated ribosomal formation, structure, and function. This review provides an overview of the current understanding of the assembly of the 30S ribosomal subunit. The 30S subunit contains one rRNA molecule (16S) and 21 ribosomal proteins (r-proteins; S1 to S21). The formation of functional subunits can occur as a self-assembly process in vitro; i.e., all the information required for the formation of active ribosomes resides in the primary sequences of the r-proteins and rRNAs. In vitro reconstitution of functional 30S subunits is carried out by using a mixture of TP30, individually purified natural or recombinant r-proteins, and natural 16S rRNA. Chemical probing and primer extension analysis have been used extensively to monitor changes in the reactivities of nucleotides in 16S rRNA during the in vitro reconstitution of 30S subunits. The potential roles for r-proteins in 30S subunit assembly were determined by omitting single proteins in reconstitution experiments. The RNPs resulting from single protein omissions were examined in terms of their composition and function to determine the roles of the absent proteins. Recent developments in understanding the structure of the 30S subunit have led to speculation about roles for some of the r-proteins in assembly. The crystal structures of the 30S subunit ( 1 , 2 ) and the 70S ribosome ( 3 ) reveal details of the r-protein and rRNA interactions.

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Figures

Image of Figure 1
Figure 1

(A) Secondary structure of 16S rRNA ( 7 , 8 ) with the major helices numbered ( 6 ). 5′, central, 3′ major, and 3′ minor domains are shown. (B) Three-dimensional structure of 16S rRNA ( 2 ) showing the 5′, central, 3′ major, and 3′ minor domains. All the three-dimensional figures were generated using the Protein Data Bank file 1J5E with ribbons ( 9 ). (C) Structure of the 30S subunit ( 2 ) showing the r-proteins that associate with the structural domains to form the body, the platform, and the head. The r-proteins that bind to specific domains are colored as in panel B. 16S rRNA is shown in grey.

Citation: Culver G, Kirthi N. 2008. Assembly of the 30S Ribosomal Subunit, EcoSal Plus 2008; doi:10.1128/ecosalplus.2.5.3
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Image of Figure 2
Figure 2

(A) Assembly map of the 30S subunit ( 15 , 18 , 19 ) showing the interaction between 16S rRNA and r-proteins. 16S rRNA is depicted as a rectangle. The arrows indicate the interaction among the ribosomal components. The dashed-line box indicates that S6 and S18 bind as a heterodimer. (B) Three-dimensional structure of the 30S subunit ( 2 ) showing the positions of primary, secondary, and tertiary proteins. 16S rRNA is shown in grey. The r-proteins are colored as in panel A.

Citation: Culver G, Kirthi N. 2008. Assembly of the 30S Ribosomal Subunit, EcoSal Plus 2008; doi:10.1128/ecosalplus.2.5.3
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Image of Figure 3
Figure 3

(A) Primary and secondary proteins, shown in red, bind to 16S rRNA, shown in grey, to form the 21S RI. (B) The RI undergoes conformational change to form RI* by heat treatment (?). (C) RI* binds to tertiary proteins, shown in blue, to form the functional 30S subunit.

Citation: Culver G, Kirthi N. 2008. Assembly of the 30S Ribosomal Subunit, EcoSal Plus 2008; doi:10.1128/ecosalplus.2.5.3
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Image of Figure 4
Figure 4

(A) Kinetic data ( 26 , 27 ) represented on the in vitro 30S subunit assembly map ( 15 , 18 , 19 ). 16S rRNA is represented as a rectangle. The early-assembly proteins are represented in red. The mid- and mid-to-late-assembly proteins are represented in yellow and green, respectively. The late-assembly proteins are shown in blue. Arrows indicate the interactions between the ribosomal components. (B) Three-dimensional structure of the 30S subunit ( 2 ) showing the relative positions of early-, mid-, mid-to-late-, and late-assembly proteins. The r-proteins are colored as in panel A, and 16S rRNA is shown in grey.

Citation: Culver G, Kirthi N. 2008. Assembly of the 30S Ribosomal Subunit, EcoSal Plus 2008; doi:10.1128/ecosalplus.2.5.3
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