A Listeria monocytogenes RNA Helicase Essential for Growth and Ribosomal Maturation at Low Temperatures Uses Its C Terminus for Appropriate Interaction with the Ribosome

Netterling, Sakura; Vaitkevicius, Karolis; Nord, Stefan; Johansson, Jörgen

doi:10.1128/jb.00348-12

Cited by 17 publications

(27 citation statements)

References 58 publications

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“…Overall, the results indicate that intact CrhR is required for proper cellular localization. This conclusion is similar to those from studies revealing similar responses to C-terminal deletion of a cold-induced RNA helicase, Lmo1722, which resulted in disassociation from the 50S ribosome in Listeria monocytogenes (38) and removal of CshA from the Staphylococcus aureus degradosome (14). While thylakoid membrane-associated polysomes have been reported in cyanobacteria (42), the subcellular localization of the cyanobacterial degradosome has not been determined (39).…”

Section: Figsupporting

confidence: 73%

“…Although many prokaryotic DEAD box RNA helicases have roles in ribosome biogenesis (16)(17)(18)38), our polysome fractionation results suggest that this function is unlikely in Synechocystis, as CrhR was detected only in the polysome-containing fraction. In addition, our runoff polysome analysis indicated that CrhR is as- a Warm-grown (30°C) and cold-shocked (20°C for 3 h) cells were viewed using immunoelectron microscopy.…”

Section: Discussionmentioning

confidence: 76%

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Cyanobacterial RNA Helicase CrhR Localizes to the Thylakoid Membrane Region and Cosediments with Degradosome and Polysome Complexes in Synechocystis sp. Strain PCC 6803

et al. 2016

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The cyanobacterium Synechocystis sp. strain PCC 6803 encodes a single DEAD box RNA helicase, CrhR, whose expression is tightly autoregulated in response to cold stress. Subcellular localization and proteomic analysis results indicate that CrhR localizes to both the cytoplasmic and thylakoid membrane regions and cosediments with polysome and RNA degradosome components. Evidence is presented that either functional RNA helicase activity or a C-terminal localization signal was required for polysome but not thylakoid membrane localization. Polysome fractionation and runoff translation analysis results indicate that CrhR associates with actively translating polysomes. The data implicate a role for CrhR in translation or RNA degradation in the thylakoid region related to thylakoid biogenesis or stability, a role that is enhanced at low temperature. Furthermore, CrhR cosedimentation with polysome and RNA degradosome complexes links alteration of RNA secondary structure with a potential translation-RNA degradation complex in Synechocystis. IMPORTANCEThe interaction between mRNA translation and degradation is a major determinant controlling gene expression. Regulation of RNA function by alteration of secondary structure by RNA helicases performs crucial roles, not only in both of these processes but also in all aspects of RNA metabolism. Here, we provide evidence that the cyanobacterial RNA helicase CrhR localizes to both the cytoplasmic and thylakoid membrane regions and cosediments with actively translating polysomes and RNA degradosome components. These findings link RNA helicase alteration of RNA secondary structure with translation and RNA degradation in prokaryotic systems and contribute to the data supporting the idea of the existence of a macromolecular machine catalyzing these reactions in prokaryotic systems, an association hitherto recognized only in archaea and eukarya. C ompartmentalization of metabolic processes is crucial for cells to avoid futile cycles. While this is efficiently achieved by lipid bilayers in eukaryotic cells, this solution does not generally apply in prokaryotes. Although prokaryotes have the ability to compartmentalize using invagination of the inner cell membrane, these compartments, for example, magnetosomes, tend to perform one specific function and are not widely used (1). Prokaryotes can also form proteinaceous microcompartments in which specific reactions occur, the primary cyanobacterial example being that of carboxysomes (2). Prokaryotes do not compartmentalize routine, everyday anabolic and catabolic processes such as transcription and translation or protein and RNA decay; however, there is a growing body of evidence indicating that prokaryotes spatially confine these cellular processes (3). A prime example is the localization of the RNA degradation complex, the degradosome, to the cytoplasmic membrane (CM) in Escherichia coli (4) and Bacillus subtilis (5).The E. coli degradosome consists of RNase E, polynucleotide phosphorylase (PNPase), enolase, and the DEAD box RNA he...

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Section: Figsupporting

confidence: 73%

Section: Discussionmentioning

confidence: 76%

Cyanobacterial RNA Helicase CrhR Localizes to the Thylakoid Membrane Region and Cosediments with Degradosome and Polysome Complexes in Synechocystis sp. Strain PCC 6803

et al. 2016

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“…Defects in ribosome biogenesis were already investigated in E. coli. There, deletion of srmB or csdA leads to a decrease in free 50S subunits and accumulation of 40S particles representing incompletely assembled large subunits (14,53). Interestingly, even though we also observe a clear reduction of the 50S subunits, no 40S premature particles were detected.…”

Section: Discussionmentioning

confidence: 65%

DEAD-Box RNA Helicases in Bacillus subtilis Have Multiple Functions and Act Independently from Each Other

Lehnik-Habrink¹,

Rempeters²,

Kovács³

et al. 2012

Journal of Bacteriology

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d DEAD-box RNA helicases play important roles in remodeling RNA molecules and in facilitating a variety of RNA-protein interactions that are key to many essential cellular processes. In spite of the importance of RNA, our knowledge about RNA helicases is limited. In this study, we investigated the role of the four DEAD-box RNA helicases in the Gram-positive model organism Bacillus subtilis. A strain deleted of all RNA helicases is able to grow at 37°C but not at lower temperatures. The deletion of cshA, cshB, or yfmL in particular leads to cold-sensitive phenotypes. Moreover, these mutant strains exhibit unique defects in ribosome biogenesis, suggesting distinct functions for the individual enzymes in this process. Based on protein accumulation, severity of the cold-sensitive phenotype, and the interaction with components of the RNA degradosome, CshA is the major RNA helicase of B. subtilis. To unravel the functions of CshA in addition to ribosome biogenesis, we conducted microarray analysis and identified the ysbAB and frlBONMD mRNAs as targets that are strongly affected by the deletion of the cshA gene. Our findings suggest that the different helicases make distinct contributions to the physiology of B. subtilis. Ribosome biogenesis and RNA degradation are two of their major tasks in B. subtilis. RNA molecules participate in a large number of cellular processes. Due to their relative simplicity (they are composed of only four different nucleic acid building blocks) and the molecular crowding found in the cytoplasm of cells, RNAs are prone to form undesired inter-and intramolecular interactions. In order to prevent these interactions, every cell expresses proteins that assist the RNA molecules to function properly. A major class of these proteins are DEAD-box RNA helicases (1).DEAD-box RNA helicases are ubiquitous enzymes that consist of a highly conserved helicase core comprising two RecA-like domains. Within the two domains, 12 characteristic sequence motifs have been identified. These motifs are involved in ATP and RNA binding, ATP hydrolysis, and the communication between the different sites. Motif II contains the name-giving Asp-Glu-AlaAsp, or DEAD, amino acid sequence (for reviews, see references 1 and 2). In addition to the conserved helicase core, most DEADbox proteins contain variable N-or C-terminal extensions. These additional domains often confer specificity for their substrates and interaction partners and are thus involved in the specific functional output and regulation of the DEAD-box protein (3,4,5,6).

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“…Involvement of RNA helicases during RNA maturation and degradation has been shown in Escherichia coli (RhlB and RhlE), Bacillus subtilis (CshA, CshB, DeaD, and YfmL), and Staphylococcus aureus (CshA) where they also been shown to associate with RNA degradosome components (14,(16)(17)(18)(19)(20)(21)(22)(23). One RNA helicase in Listeria monocytogenes, Lmo1722, was shown to be important for ribosome biogenesis and interacted with the 50S subunit through its C terminus (6). Not surprisingly, the absence of Lmo1722 and other listerial RNA helicases has the largest effect at low temperatures and under adverse stress conditions (6,15,(24)(25)(26)(27).…”

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confidence: 99%

“…One RNA helicase in Listeria monocytogenes, Lmo1722, was shown to be important for ribosome biogenesis and interacted with the 50S subunit through its C terminus (6). Not surprisingly, the absence of Lmo1722 and other listerial RNA helicases has the largest effect at low temperatures and under adverse stress conditions (6,15,(24)(25)(26)(27).…”

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confidence: 99%

RNA Helicase Important for Listeria monocytogenes Hemolytic Activity and Virulence Factor Expression

et al. 2016

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RNA helicases have been shown to be important for the function of RNA molecules at several levels, although their putative involvement in microbial pathogenesis has remained elusive. We have previously shown that Listeria monocytogenes DExD-box RNA helicases are important for bacterial growth, motility, ribosomal maturation, and rRNA processing. We assessed the importance of the RNA helicase Lmo0866 (here named CshA) for expression of virulence traits. We observed a reduction in hemolytic activity in a strain lacking CshA compared to the wild type. This phenomenon was less evident in strains lacking other RNA helicases. The reduced hemolysis was accompanied by lower expression of major listerial virulence factors in the ΔcshA strain, mainly listeriolysin O, but also to some degree the actin polymerizing factor ActA. Reduced expression of these virulence factors in the strain lacking CshA did not, however, correlate with a decreased level of the virulence regulator PrfA. When combining the ΔcshA knockout with a mutation creating a constitutively active PrfA protein (PrfA*), the effect of the ΔcshA knockout on LLO expression was negated. These data suggest a role for the RNA helicase CshA in posttranslational activation of PrfA. Surprisingly, although the expression of several virulence factors was reduced, the ΔcshA strain did not demonstrate any reduced ability to infect nonphagocytic cells compared to the wild-type strain.

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A Listeria monocytogenes RNA Helicase Essential for Growth and Ribosomal Maturation at Low Temperatures Uses Its C Terminus for Appropriate Interaction with the Ribosome

Cited by 17 publications

References 58 publications

Cyanobacterial RNA Helicase CrhR Localizes to the Thylakoid Membrane Region and Cosediments with Degradosome and Polysome Complexes in Synechocystis sp. Strain PCC 6803

Cyanobacterial RNA Helicase CrhR Localizes to the Thylakoid Membrane Region and Cosediments with Degradosome and Polysome Complexes in Synechocystis sp. Strain PCC 6803

DEAD-Box RNA Helicases in Bacillus subtilis Have Multiple Functions and Act Independently from Each Other

RNA Helicase Important for Listeria monocytogenes Hemolytic Activity and Virulence Factor Expression

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