A Listeria monocytogenes RNA helicase essential for growth and ribosomal maturation at low temperatures uses its C terminus for appropriate interaction with the ribosome.
Listeria monocytogenes, a Gram-positive food-borne human pathogen, is able to grow at temperatures close to 0°C and is thus of great concern for the food industry. In this work, we investigated the physiological role of one DExD-box RNA helicase in Listeria monocytogenes. The RNA helicase Lmo1722 was required for optimal growth at low temperatures, whereas it was dispensable at 37°C. A Δlmo1722 strain was less motile due to downregulation of the major subunit of the flagellum, FlaA, caused by decreased flaA expression. By ribosomal fractionation experiments, it was observed that Lmo1722 was mainly associated with the 50S subunit of the ribosome. Absence of Lmo1722 decreased the fraction of 50S ribosomal subunits and mature 70S ribosomes and affected the processing of the 23S precursor rRNA. The ribosomal profile could be restored to wild-type levels in a Δlmo1722 strain expressing Lmo1722. Interestingly, the C-terminal part of Lmo1722 was redundant for low-temperature growth, motility, 23S rRNA processing, and appropriate ribosomal maturation. However, Lmo1722 lacking the C terminus showed a reduced affinity for the 50S and 70S fractions, suggesting that the C terminus is important for proper guidance of Lmo1722 to the 50S subunit. Taken together, our results show that the Listeria RNA helicase Lmo1722 is essential for growth at low temperatures, motility, and rRNA processing and is important for ribosomal maturation, being associated mainly with the 50S subunit of the ribosome.
Department of Molecular Biology, Umeå University, Umeå, Sweden.
Sakura
S
Netterling
Karolis
K
Vaitkevicius
Stefan
S
Nord
Jörgen
J
Johansson
D017353:Gene Deletion; D005816:Genetic Complementation Test; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000254:growth & development* / Q000378:metabolism; D008124:Locomotion; D011485:Protein Binding; D025941:Protein Interaction Mapping; D020365:RNA Helicases / Q000235:genetics / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012338:RNA, Ribosomal, 23S / Q000378:metabolism*; D013696:Temperature
D012338:RNA, Ribosomal, 23S; D020365:RNA Helicases