Article: Demonstration of the role of the DnaK chaperone system in assembly of 30S ribosomal subunits using a purified in vitro system

Recently, there has been controversy regarding the ability of the DnaK chaperone system to facilitate Escherichia coli 30S subunit assembly at otherwise nonpermissive conditions. Here, we present additional data indicating that purified DnaK chaperone assembled 30S subunits are functional. Additionally, explanations for reported differences are discussed.

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References

1.

El Hage A, Sbai M, Alix J . The chaperonin GroEL and other heat-shock proteins, besides DnaK, participate in ribosome biogenesis in Escherichia coli. Mol Gen Genet. 2001; 264(6):796-808. DOI: 10.1007/s004380000369. View

2.

Maki J, Schnobrich D, Culver G . The DnaK chaperone system facilitates 30S ribosomal subunit assembly. Mol Cell. 2002; 10(1):129-38. DOI: 10.1016/s1097-2765(02)00562-2. View

3.

Southworth D, Brunelle J, Green R . EFG-independent translocation of the mRNA:tRNA complex is promoted by modification of the ribosome with thiol-specific reagents. J Mol Biol. 2002; 324(4):611-23. DOI: 10.1016/s0022-2836(02)01196-8. View

4.

Williamson J . After the ribosome structures: how are the subunits assembled?. RNA. 2003; 9(2):165-7. PMC: 1370380. DOI: 10.1261/rna.2164903. View

5.

Charollais J, Pflieger D, Vinh J, Dreyfus M, Iost I . The DEAD-box RNA helicase SrmB is involved in the assembly of 50S ribosomal subunits in Escherichia coli. Mol Microbiol. 2003; 48(5):1253-65. DOI: 10.1046/j.1365-2958.2003.03513.x. View

6.

Culver G, Noller H . Efficient reconstitution of functional Escherichia coli 30S ribosomal subunits from a complete set of recombinant small subunit ribosomal proteins. RNA. 1999; 5(6):832-43. PMC: 1369808. DOI: 10.1017/s1355838299990714. View

7.

Traub P, Nomura M . Structure and function of E. coli ribosomes. V. Reconstitution of functionally active 30S ribosomal particles from RNA and proteins. Proc Natl Acad Sci U S A. 1968; 59(3):777-84. PMC: 224743. DOI: 10.1073/pnas.59.3.777. View

8.

Held W, Nomura M . Rate determining step in the reconstitution of Escherichia coli 30S ribosomal subunits. Biochemistry. 1973; 12(17):3273-81. DOI: 10.1021/bi00741a020. View

9.

Paek K, Walker G . Escherichia coli dnaK null mutants are inviable at high temperature. J Bacteriol. 1987; 169(1):283-90. PMC: 211765. DOI: 10.1128/jb.169.1.283-290.1987. View

10.

Alix J, Guerin M . Mutant DnaK chaperones cause ribosome assembly defects in Escherichia coli. Proc Natl Acad Sci U S A. 1993; 90(20):9725-9. PMC: 47643. DOI: 10.1073/pnas.90.20.9725. View

11.

Alix J, Nierhaus K . DnaK-facilitated ribosome assembly in Escherichia coli revisited. RNA. 2003; 9(7):787-93. PMC: 1370445. DOI: 10.1261/rna.5360203. View

Demonstration of the Role of the DnaK Chaperone System in Assembly of 30S Ribosomal Subunits Using a Purified in Vitro System