Journal article

Publication year: 2005

Pages: 14572-14578

Journal: Journal of Biological Chemistry

Volume number: 280

Issue number: 15

ISSN: 0021-9258

Open access status: Hybrid

DOI Link: https://doi.org/10.1074/jbc.M413507200

Publisher: Elsevier

Abstract: 
Endoribonuclease E, a key enzyme involved in RNA decay and processing in bacteria, organizes a protein complex called degradosome. In Escherichia coli, Rhodobacter capsulatus, and Streptomyces coelicolor, RNase E interacts with the phosphate-dependent exoribonuclease polynucleotide phosphorylase, DEAD-box helicase(s), and additional factors in an RNA-degrading complex. To characterize the degradosome of the psychrotrophic bacterium Pseudomonas syringae Lz4W, RNase E was enriched by cation exchange chromatography and fractionation in a glycerol density gradient. Most surprisingly, the hydrolytic exoribonuclease RNase R was found to co-purify with RNase E. Co-immunoprecipitation and Ni2+-affinity pull-down experiments confirmed the specific interaction between RNase R and RNase E. Additionally, the DEAD-box helicase RhlE was identified as part of this protein complex. Fractions comprising the three proteins showed RNase E and RNase R activity and efficiently degraded a synthetic stem-loop containing RNA in the presence of ATP. The unexpected association of RNase R with RNase E and RhlE in an RNA-degrading complex indicates that the cold-adapted P. syringae has a degradosome of novel structure. The identification of RNase R instead of polynucleotide phosphorylase in this complex underlines the importance of the interaction between endo- and exoribonucleases for the bacterial RNA metabolism. The physical association of RNase E with an exoribonuclease and an RNA helicase apparently is a common theme in the composition of bacterial RNA-degrading complexes.

Harvard Citation style: Purusharth, R., Klein, F., Sulthana, S., Jager, S., Jagannadham, M., Evguenieva-Hackenberg, E., et al. (2005) Exoribonuclease R interacts with endoribonuclease e and an RNA helicase in the psychrotrophic Bacterium pseudomonas syringae Lz4W, Journal of Biological Chemistry, 280(15), pp. 14572-14578. https://doi.org/10.1074/jbc.M413507200

APA Citation style: Purusharth, R., Klein, F., Sulthana, S., Jager, S., Jagannadham, M., Evguenieva-Hackenberg, E., Ray, M., & Klug, G. (2005). Exoribonuclease R interacts with endoribonuclease e and an RNA helicase in the psychrotrophic Bacterium pseudomonas syringae Lz4W. Journal of Biological Chemistry. 280(15), 14572-14578. https://doi.org/10.1074/jbc.M413507200