Ribonucleoprotein formation by the ORF1 protein of the non-LTR retrotransposon Tx1L in Xenopus oocytes.
The Tx1L elements constitute a family of site-specific non-LTR retrotransposons found in the genome of the frog Xenopus laevis . The elements have two open reading frames (ORFs) with homology to proteins of retroviruses and other retroelements. This study demonstrates an expected activity of one of the element-encoded proteins. The RNA binding properties of ORF1p, the product of the first ORF of Tx1L, were examined after expression from RNA injected into Xenopus oocytes. Using sucrose gradient sedimentation and non-denaturing gel electrophoresis, we show that ORF1p associates with RNA in cytoplasmic ribonucleoprotein (RNP) particles. Discrete RNPs are formed with well-defined mobilities. The ORF1p RNPs are distinct from endogenous RNPs that contain stored oocyte mRNAs and two specific endogenous mRNAs do not become associated with ORF1p. ORF1p appears to be capable of associating with its own mRNA and with other injected RNAs, independent of specific recognition sequences. Although nuclear localization of ORF1p was anticipated, based both on the supposed mechanism of transposition and on the presence of a potential nuclear localization signal, no significant fraction of the protein was found in the oocyte nucleus. Nonetheless, the RNA binding capability of ORF1p is consistent with the proposed model for transposition of non-LTR retrotransposons.
Pont-Kingdon
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G
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84132, USA. genevieve.kingdon@genetics.utah.edu
Chi
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Christensen
S
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Carroll
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D000818:Animals; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D046911:Macromolecular Substances; D009865:Oocytes; D016366:Open Reading Frames; D012313:RNA / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D018626:Retroelements; D012261:Ribonucleoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis
D046911:Macromolecular Substances; D018626:Retroelements; D012261:Ribonucleoproteins; D012313:RNA