A second RNA-binding protein is essential for ethanol tolerance provided by the bacterial OLE ribonucleoprotein complex

OLE (ornate, large, extremophilic) RNAs comprise a class of structured noncoding RNAs (ncRNAs) found in many extremophilic bacteria species. OLE RNAs constitute one of the longest and most widespread bacterial ncRNA classes whose major biochemical function remains unknown. In the Gram-positive alkaliphile Bacillus halodurans, OLE RNA is abundant, and localizes to the cell membrane by association with the transmembrane OLE-associated protein called OapA (formerly OAP). These characteristics, along with the well-conserved sequence and structural features of OLE RNAs, suggest that the OLE ribonucleoprotein (RNP) complex performs important biological functions. B. halodurans strains lacking OLE RNA (∆ole) or OapA (∆oapA) are less tolerant of cold (20 °C) and short-chain alcohols (e.g., ethanol). Here, we describe the effects of a mutant OapA (called PM1) that more strongly inhibits growth under cold or ethanol stress compared with strains lacking the oapA gene, even when wild-type OapA is present. This dominant-negative effect of PM1 is reversed by mutations that render OLE RNA nonfunctional. This finding demonstrates that the deleterious PM1 phenotype requires an intact RNP complex, and suggests that the complex has one or more additional undiscovered components. A genetic screen uncovered PM1 phenotype suppressor mutations in the ybzG gene, which codes for a putative RNA-binding protein of unknown biological function. We observe that YbzG protein (also called OapB) selectively binds OLE RNA in vitro, whereas a mutant version of the protein is not observed to bind OLE RNA. Thus, YbzG/OapB is an important component of the functional OLE RNP complex in B. halodurans.

Download Full-text

GPKOW is essential for pre-mRNA splicing in vitro and suppresses splicing defect caused by dominant-negative DHX16 mutation in vivo

Bioscience Reports ◽

10.1042/bsr20140142 ◽

2014 ◽

Vol 34 (6) ◽

Cited By ~ 7

Author(s):

Shengbing Zang ◽

Ting-Yu Lin ◽

Xinji Chen ◽

Marieta Gencheva ◽

Alain N. S. Newo ◽

...

Keyword(s):

Rna Splicing ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Mrna Splicing ◽

Dominant Negative ◽

Splicing Defect ◽

Similarity And Difference

Using biochemical, mutation, and cellular analyses, we characterized important domains involved in the functionality of a RNA-binding protein in RNA splicing. We also showed the similarity and difference between yeast and human counterparts.

Download Full-text

Cloning and sequencing of the cDNA for an RNA-binding protein from the Mexican axolotl: binding affinity of the in vitro synthesized protein

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression ◽

10.1016/s0167-4781(98)00071-2 ◽

1998 ◽

Vol 1398 (3) ◽

pp. 265-274 ◽

Cited By ~ 1

Author(s):

Rajula Bhatia ◽

Arun Gaur ◽

Larry F. Lemanski ◽

Dipak K. Dube

Keyword(s):

Binding Affinity ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Cloning And Sequencing ◽

Mexican Axolotl

Download Full-text

Characterization and purification of lupus antigen La, and RNA-binding protein

Molecular and Cellular Biology ◽

10.1128/mcb.5.3.586-590.1985 ◽

1985 ◽

Vol 5 (3) ◽

pp. 586-590

Author(s):

A M Francoeur ◽

E K Chan ◽

J I Garrels ◽

M B Mathews

Keyword(s):

Hela Cell ◽

Gel Electrophoresis ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

In Vitro Translation ◽

Cell Protein ◽

Two Dimensional Gel Electrophoresis ◽

La Antigen

HeLa cell La antigen, an RNA-binding protein, was characterized by using two-dimensional gel electrophoresis. Eight isoelectric forms (pI 6 to 7) were observed, many containing phosphate. An in vitro translation product similar in size and antigenicity was identified. The HeLa cell protein purified by using an assay based on ribonucleoprotein reconstitution with adenovirus VA RNAI also comprised several isoelectric forms.

Download Full-text

The viral nucleocapsid protein and the human RNA-binding protein Mex3A promote translation of the Andes orthohantavirus small mRNA

PLoS Pathogens ◽

10.1371/journal.ppat.1009931 ◽

2021 ◽

Vol 17 (9) ◽

pp. e1009931

Author(s):

Jorge Vera-Otarola ◽

Estefania Castillo-Vargas ◽

Jenniffer Angulo ◽

Francisco M. Barriga ◽

Eduard Batlle ◽

...

Keyword(s):

Nucleocapsid Protein ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Initiation Factor ◽

Cellular Proteins ◽

Dependent Manner ◽

The Andes ◽

Viral Nucleocapsid

The capped Small segment mRNA (SmRNA) of the Andes orthohantavirus (ANDV) lacks a poly(A) tail. In this study, we characterize the mechanism driving ANDV-SmRNA translation. Results show that the ANDV-nucleocapsid protein (ANDV-N) promotes in vitro translation from capped mRNAs without replacing eukaryotic initiation factor (eIF) 4G. Using an RNA affinity chromatography approach followed by mass spectrometry, we identify the human RNA chaperone Mex3A (hMex3A) as a SmRNA-3’UTR binding protein. Results show that hMex3A enhances SmRNA translation in a 3’UTR dependent manner, either alone or when co-expressed with the ANDV-N. The ANDV-N and hMex3A proteins do not interact in cells, but both proteins interact with eIF4G. The hMex3A–eIF4G interaction showed to be independent of ANDV-infection or ANDV-N expression. Together, our observations suggest that translation of the ANDV SmRNA is enhanced by a 5’-3’ end interaction, mediated by both viral and cellular proteins.

Download Full-text

Regulation of cold-induced thermogenesis by the RNA binding protein FAM195A

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2104650118 ◽

2021 ◽

Vol 118 (23) ◽

pp. e2104650118

Author(s):

Jessica Cannavino ◽

Mengle Shao ◽

Yu A. An ◽

Svetlana Bezprozvannaya ◽

Shiuhwei Chen ◽

...

Keyword(s):

Rna Binding ◽

Binding Protein ◽

Uncoupling Protein ◽

Rna Binding Protein ◽

Cold Environments ◽

Transport Chain ◽

Brown Adipose ◽

Mitochondrial Oxidation ◽

Branched Chain

Homeothermic vertebrates produce heat in cold environments through thermogenesis, in which brown adipose tissue (BAT) increases mitochondrial oxidation along with uncoupling of the electron transport chain and activation of uncoupling protein 1 (UCP1). Although the transcription factors regulating the expression of UCP1 and nutrient oxidation genes have been extensively studied, only a few other proteins essential for BAT function have been identified. We describe the discovery of FAM195A, a BAT-enriched RNA binding protein, which is required for cold-dependent thermogenesis in mice. FAM195A knockout (KO) mice display whitening of BAT and an inability to thermoregulate. In BAT of FAM195A KO mice, enzymes involved in branched-chain amino acid (BCAA) metabolism are down-regulated, impairing their response to cold. Knockdown of FAM195A in brown adipocytes in vitro also impairs expression of leucine oxidation enzymes, revealing FAM195A to be a regulator of BCAA metabolism and a potential target for metabolic disorders.

Download Full-text

Heterogeneous Nuclear Ribonucleoprotein (hnRNP) E1 Binds to hnRNP A2 and Inhibits Translation of A2 Response Element mRNAs

Molecular Biology of the Cell ◽

10.1091/mbc.e05-10-0946 ◽

2006 ◽

Vol 17 (8) ◽

pp. 3521-3533 ◽

Cited By ~ 51

Author(s):

Linda D. Kosturko ◽

Michael J. Maggipinto ◽

George Korza ◽

Joo Won Lee ◽

John H. Carson ◽

...

Keyword(s):

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

In Vitro Translation ◽

Dependent Manner ◽

Response Element ◽

Heterogeneous Nuclear Ribonucleoprotein ◽

Nuclear Ribonucleoprotein ◽

Hnrnp E1

Heterogeneous nuclear ribonucleoprotein (hnRNP) A2 is a trans-acting RNA-binding protein that mediates trafficking of RNAs containing the cis-acting A2 response element (A2RE). Previous work has shown that A2RE RNAs are transported to myelin in oligodendrocytes and to dendrites in neurons. hnRNP E1 is an RNA-binding protein that regulates translation of specific mRNAs. Here, we show by yeast two-hybrid analysis, in vivo and in vitro coimmunoprecipitation, in vitro cross-linking, and fluorescence correlation spectroscopy that hnRNP E1 binds to hnRNP A2 and is recruited to A2RE RNA in an hnRNP A2-dependent manner. hnRNP E1 is colocalized with hnRNP A2 and A2RE mRNA in granules in dendrites of oligodendrocytes. Overexpression of hnRNP E1 or microinjection of exogenous hnRNP E1 in neural cells inhibits translation of A2RE mRNA, but not of non-A2RE RNA. Excess hnRNP E1 added to an in vitro translation system reduces translation efficiency of A2RE mRNA, but not of nonA2RE RNA, in an hnRNP A2-dependent manner. These results are consistent with a model where hnRNP E1 recruited to A2RE RNA granules by binding to hnRNP A2 inhibits translation of A2RE RNA during granule transport.

Download Full-text

Involvement of a site-specific trans-acting factor and a common RNA-binding protein in the editing of chloroplast mRNAs: development of a chloroplast in vitro RNA editing system

The EMBO Journal ◽

10.1093/emboj/20.5.1144 ◽

2001 ◽

Vol 20 (5) ◽

pp. 1144-1152 ◽

Cited By ~ 109

Author(s):

T. Hirose

Keyword(s):

Rna Editing ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Site Specific ◽

A Site

Download Full-text

The RNA-Binding Protein Hfq of Listeria monocytogenes: Role in Stress Tolerance and Virulence

Journal of Bacteriology ◽

10.1128/jb.186.11.3355-3362.2004 ◽

2004 ◽

Vol 186 (11) ◽

pp. 3355-3362 ◽

Cited By ~ 176

Author(s):

Janne K. Christiansen ◽

Marianne H. Larsen ◽

Hanne Ingmer ◽

Lotte Søgaard-Andersen ◽

Birgitte H. Kallipolitis

Keyword(s):

Listeria Monocytogenes ◽

Stress Tolerance ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Stationary Growth Phase ◽

Ethanol Stress ◽

Gram Positive ◽

Term Survival

ABSTRACT In gram-negative bacteria, the RNA-binding protein Hfq has emerged as an important regulatory factor in a variety of physiological processes, including stress resistance and virulence. In Escherichia coli, Hfq modulates the stability or the translation of mRNAs and interacts with numerous small regulatory RNAs. Here, we studied the role of Hfq in the stress tolerance and virulence of the gram-positive food-borne human pathogen Listeria monocytogenes. We present evidence that Hfq is involved in the ability of L. monocytogenes to tolerate osmotic and ethanol stress and contributes to long-term survival under amino acid-limiting conditions. However, Hfq is not required for resistance to acid and oxidative stress. Transcription of hfq is induced under various stress conditions, including osmotic and ethanol stress and at the entry into the stationary growth phase, thus supporting the view that Hfq is important for the growth and survival of L. monocytogenes in harsh environments. The stress-inducible transcription of hfq depends on the alternative sigma factor σB, which controls the expression of numerous stress- and virulence-associated genes in L. monocytogenes. Infection studies showed that Hfq contributes to pathogenesis in mice, yet plays no role in the infection of cultured cell lines. This study provides, for the first time, information on the role of Hfq in the stress tolerance and virulence of a gram-positive pathogen.

Download Full-text