The pseudouridine synthase RluD is required for normal ribosome assembly and function in Escherichia coli

RNA - Tập 11 Số 7 - Trang 1141-1152 - 2005
N S Gutgsell1, Murray P. Deutscher2, James Ofengand3
1Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Gautier Bldg., 1011 NW 15th St., Miami, FL 33136, USA. [email protected]
2Biochemistry & Molecular Biology,
3University of Miami

Tóm tắt

RluD is the pseudouridine synthase responsible for the formation of Ψ1911, Ψ1915, and Ψ1917 in Escherichia coli 23S rRNA. Previous work from our laboratory demonstrated that disruption of the rluD gene and/or loss of the pseudouridine residues for which it is responsible resulted in a severe growth phenotype. In the current work we have examined further the effect of the loss of the RluD protein and its product pseudouridine residues in a deletion strain lacking the rluD gene. This strain exhibits defects in ribosome assembly, biogenesis, and function. Specifically, there is a deficit of 70S ribosomes, an increase in 50S and 30S subunits, and the appearance of new 62S and 39S particles. Analysis of the 39S particles indicates that they are immature precursors of the 50S subunits, whereas the 62S particles are derived from the breakdown of unstable 70S ribosomes. In addition, purified mutant 70S ribosomes were found to be somewhat less efficient than wild type in protein synthesis. The defect in ribosome assembly and resulting growth phenotype of the mutant could be restored by expression of wild-type RluD and synthesis of Ψ1911, Ψ1915, and Ψ1917 residues, but not by catalytically inactive mutant RluD proteins, incapable of pseudouridine formation. The data suggest that the loss of the pseudouridine residues can account for all aspects of the mutant phenotype; however, a possible second function of the RluD synthase is also discussed.

Từ khóa


Tài liệu tham khảo

10.1073/pnas.0401904101

1976, Bacteriol. Rev., 40, 116, 10.1128/br.40.1.116-167.1976

10.1016/S1097-2765(03)00009-1

10.1074/jbc.M409581200

10.1046/j.1365-2958.2003.03513.x

10.1093/nar/gkh603

10.1074/jbc.273.29.18562

10.1017/S1355838299990167

10.1021/bi00163a008

10.1073/pnas.120163297

2001, RNA, 7, 1603

10.1261/rna.5187404

10.1073/pnas.81.14.4290

10.1046/j.1365-2958.2003.03813.x

10.1128/JB.182.17.4719-4729.2000

10.1038/71219

10.1017/S1355838200001588

10.1017/S1355838201000243

10.1074/jbc.M205423200

10.1261/rna.5230603

10.1074/jbc.M309324200

10.1017/S1355838299981669

10.1016/0022-2836(75)90089-3

10.1261/rna.5112704

Miller, J.H. 1992. A short course in bacterial genetics: A laboratory manual and handbook for Escherichia coli and related bacteria, Chapter 28, pp. 357–364. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

1979, Methods Enzymol., LIX, 443

Ofengand, J. and Rudd, K.E. 2000. The ribosome: Structure, function, antibiotics and cellular interactions. In The ribosome: Structure, function, antibiotics, and cellular interactions (eds. R.A. Garrett et al.), pp. 175–189. ASM Press, Washington, DC.

10.1006/meth.2001.1249

10.1073/pnas.89.9.3995

10.1017/S1355838298981146

10.1074/jbc.274.27.18880

Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989. Molecular cloning: A laboratory manual, 2d ed., pp, 1.82–1.83. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

10.1093/nar/13.14.5041

10.1073/pnas.85.19.7144

10.1146/annurev.mi.44.100190.000541

10.1093/nar/18.12.3515

1971, J. Bacteriol., 107, 87, 10.1128/jb.107.1.87-94.1971

10.1146/annurev.micro.58.030603.123822

10.1126/science.1060089

Thông tin
Thông tin xuất bản

RNA

Tập 11 Số 7

1141-1152

Thông tin tác giả