Ribosome-bound Mrna

Ribosome-bound mRNA is a messenger RNA molecule associated with one or more ribosomes, where its nucleotide sequence directs the production of a specific protein. During translation, the ribosome moves along the mRNA in the 5′-to-3′ direction, reads codons, and matches them with transfer RNAs that deliver amino acids to a growing polypeptide chain. Ribosome binding can regulate when and how efficiently genes are expressed, while the formation of polysomes enables multiple ribosomes to translate one mRNA simultaneously. Studying these complexes helps explain protein synthesis, gene regulation, cellular responses, and defects linked to altered translation.

Ribosome-bound Mrna - Related Videos

Research

JoVE Journal - Biology

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

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Cited by 2 •

2012

A technique to identify translational pause sites on mRNA is described. This procedure is based on isolation of nascent polypeptides accumulating on ribosomes during in vitro translation of a target mRNA, followed by the size analysis of the nascent chains using a denaturing gel electrophoresis.

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

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Cited by 6 •

2012

We describe here a technique that is now routinely used to isolate stably bound ribosome nascent chain complexes (RNCs). This technique takes advantage of the discovery that a 17 amino acid long SecM "arrest sequence" can halt translation elongation in a prokaryotic (E. coli) system, when inserted into (or fused to the C-terminus) of virtually any protein.

Education

JoVE Core - Molecular Biology

Nonsense-mediated mRNA Decay

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2020

The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs. Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...

Regulated mRNA Transport

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2020

In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...

Research

JoVE Journal - Biology
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Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

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Cited by 2 •

2011

A major impediment to biochemical analyses of ribosomes containing nascent peptidyl-tRNAs has been the presence of other ribosomes in the same samples, ribosomes not involved in the translation of the specific mRNA sequence being analyzed. We developed a simple methodology to purify, exclusively, the ribosomes containing the nascent peptidyl-tRNA of interest.

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