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Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes

ABSTRACT: Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes

PROVIDER: PRJNA1116211 | ENA |

REPOSITORIES: ENA

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Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (Ribo-seq)

Project description:The CCR4-NOT complex is a major regulator of eukaryotic messenger RNA (mRNA) stability. Slow decoding during translation promotes association of CCR4-NOT with ribosomes, accelerating mRNA degradation. We applied selective ribosome profiling to further investigate the determinants of CCR4-NOT recruitment to ribosomes in mammalian cells. This revealed that specific arginine codons in the P-site are strong signals for ribosomal recruitment of human CNOT3, a CCR4-NOT subunit. Cryo–electron microscopy and transfer RNA (tRNA) mutagenesis demonstrated that the D-arms of select arginine tRNAs interact with CNOT3 and promote its recruitment whereas other tRNA D-arms sterically clash with CNOT3. These effects link codon content to mRNA stability. Thus, in addition to their canonical decoding function, tRNAs directly engage regulatory complexes during translation, a mechanism we term P-site tRNA-mediated mRNA decay.

2024-11-21 | GSE268324 | GEO

Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (SLAM-Seq)

2024-11-21 | GSE268322 | GEO

Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (SLAM-Seq)

Project description:Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (SLAM-Seq)

| PRJNA1116215 | ENA

Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (Ribo-seq)

Project description:Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes (Ribo-seq)

| PRJNA1116213 | ENA

Specific tRNAs promote mRNA decay by recruiting the CCR4-NOT complex to translating ribosomes

Project description:This SuperSeries is composed of the SubSeries listed below.

2024-11-21 | GSE268325 | GEO

The Ccr4-Not complex monitors the translating ribosome for codon optimality

Project description:The Ccr4-Not complex monitors the translating ribosome for codon optimality

| PRJNA543493 | ENA

The Ccr4-Not complex monitors the translating ribosome for codon optimality

Project description:The Ccr4-Not complex monitors the translating ribosome for codon optimality

| PRJNA603124 | ENA

Selective ribosome profiling to study interactions of translating ribosomes in yeast

Project description:Selective ribosome profiling to study interactions of translating ribosomes in yeast

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Specific recognition and ubiquitination of translating ribosomes by mammalian CCR4-NOT.

Project description:Translation affects messenger RNA stability and, in yeast, this is mediated by the Ccr4-Not deadenylation complex. The details of this process in mammals remain unclear. Here, we use cryogenic electron microscopy (cryo-EM) and crosslinking mass spectrometry to show that mammalian CCR4-NOT specifically recognizes ribosomes that are stalled during translation elongation in an in vitro reconstituted system with rabbit and human components. Similar to yeast, mammalian CCR4-NOT inserts a helical bundle of its CNOT3 subunit into the empty E site of the ribosome. Our cryo-EM structure shows that CNOT3 also locks the L1 stalk in an open conformation to inhibit further translation. CCR4-NOT is required for stable association of the nonconstitutive subunit CNOT4, which ubiquitinates the ribosome, likely to signal stalled translation elongation. Overall, our work shows that human CCR4-NOT not only detects but also enforces ribosomal stalling to couple translation and mRNA decay.

| S-EPMC7615087 | biostudies-literature

The Ccr4-Not complex monitors the translating ribosome for codon optimality

Project description:Control of messenger RNA (mRNA) decay rate is intimately connected to translation elongation, but the spatial coordination of these events is poorly understood. The Ccr4-Not complex initiates mRNA decay through deadenylation and activation of decapping. We used a combination of cryo–electron microscopy, ribosome profiling, and mRNA stability assays to examine the recruitment of Ccr4-Not to the ribosome via specific interaction of the Not5 subunit with the ribosomal E-site in Saccharomyces cerevisiae. This interaction occurred when the ribosome lacked accommodated A-site transfer RNA, indicative of low codon optimality. Loss of the interaction resulted in the inability of the mRNA degradation machinery to sense codon optimality. Our findings elucidate a physical link between the Ccr4-Not complex and the ribosome and provide mechanistic insight into the coupling of decoding efficiency with mRNA stability.

2020-04-16 | GSE131408 | GEO

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