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

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.

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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.

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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

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A cDNA clone for a polyadenylated RNA-binding protein of Xenopus laevis oocytes hybridizes to four developmentally regulated mRNAs

Molecular and Cellular Biology ◽

10.1128/mcb.5.10.2697-2704.1985 ◽

1985 ◽

Vol 5 (10) ◽

pp. 2697-2704

Author(s):

L J Lorenz ◽

J D Richter

Keyword(s):

Xenopus Laevis ◽

Cdna Clone ◽

Rna Binding ◽

Binding Protein ◽

Single Gene ◽

In Vitro Translation ◽

Xenopus Laevis Oocytes ◽

Cdna Insert ◽

Alternative Processing

Xenopus laevis oocytes contain a unique group of proteins which decrease during oogenesis, bind poly(A) RNA, and possibly play a role in the regulation of translation. A monoclonal antibody generated against one of these proteins was used to screen an expression vector cDNA library. A cDNA clone was isolated and confirmed to code for the binding protein by in vitro translation of hybrid-selected RNA followed by immunoprecipitation. This cDNA, when used in RNA gel blots, hybridized to four transcripts of 2.0, 1.7 (two transcripts of similar size), and 1.2 kilobases. All of the transcripts decreased in amount during oogenesis and were not evident in somatic cells. In addition, the fraction of the transcripts associated with polysomes decreased during oogenesis. Digestion of the cDNA insert with PstI generated two fragments of 220 and 480 base pairs which, when used as probes in an RNA gel blot, hybridized to unique as well as common transcripts. Genomic Southern blots suggested the presence of a single gene, indicating that these transcripts arose by alternative processing.

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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.

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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.

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