Widespread Cotranslational Formation of Protein Complexes

Duncan, Caia D. S.; Mata, Juan

doi:10.1371/journal.pgen.1002398

Cited by 112 publications

(135 citation statements)

References 37 publications

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“…Previous studies in yeast have demonstrated that transcripts associated with specific cellular functions can be copurified with one or more of the encoded proteins (26,27). Current models Fig.…”

Section: Discussionmentioning

confidence: 99%

“…suggest the transcripts are coimmunoprecipitated as a consequence of cotranslational assembly of their encoded proteins. For example, tip1 and tea2p transcripts encoding components of a molecular motor could be copurified with the Tea2p kinesin protein, but not when the tip1 translation initiation site was mutated; thus, the association occurs via the nascent proteins and not directly through the transcripts (26). A similar model was suggested by studies on the SET1C histone methyltransferase complex, which were among the first studies demonstrating that the association of protein with a transcript was not through binding of the protein to the mRNA but rather by copurification of the polysomal complex comprising the transcript and the nascent protein emerging from the ribosome (28).…”

Section: Discussionmentioning

confidence: 99%

“…The mRNAs encoding proteins in macromolecular complexes mediating specific cellular functions, such as cell cycle control, were also copurified with mRNAs encoding associated RNABPs (26). In mammalian systems, RNABPs have been characterized in neurons, where they are associated with "biologically coherent" (34) groups of mRNA encoding proteins related to a recognized function [e.g., synaptic transmission (35-37)].…”

Section: Discussionmentioning

confidence: 99%

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Cotranslational association of mRNA encoding subunits of heteromeric ion channels

Liu

Jones

Lange

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Oligomers of homomeric voltage-gated potassium channels associate early in biogenesis as the nascent proteins emerge from the polysome. Less is known about how proteins emerging from different polysomes associate to form hetero-oligomeric channels. Here, we report that alternate mRNA transcripts encoding human ether-à-go-go-related gene (hERG) 1a and 1b subunits, which assemble to produce ion channels mediating cardiac repolarization, are physically associated during translation. We show that shRNA specifically targeting either hERG 1a or 1b transcripts reduced levels of both transcripts, but only when they were coexpressed heterologously. Both transcripts could be copurified with an Ab against the nascent hERG 1a N terminus. This interaction occurred even when translation of 1b was prevented, indicating the transcripts associate independent of their encoded proteins. The association was also demonstrated in cardiomyocytes, where levels of both hERG transcripts were reduced by either 1a or 1b shRNA, but native KCNE1 and ryanodine receptor 2 (RYR2) transcripts were unaffected. Changes in protein levels and membrane currents mirrored changes in transcript levels, indicating the targeted transcripts were undergoing translation. The physical association of transcripts encoding different subunits provides the spatial proximity required for nascent proteins to interact during biogenesis, and may represent a general mechanism facilitating assembly of heteromeric protein complexes involved in a range of biological processes.KCNH2 | hERG | cotranslational assembly | microtranslatome | hERG 1a/1b

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Cotranslational association of mRNA encoding subunits of heteromeric ion channels

Liu

Jones

Lange

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…Considering the evidence from fission yeast 16 and bacteria 17 that many, if not most, multimeric protein complexes assemble co-translationally, localized translation could represent a spatial component of this "window," and indeed septins in other fungi display clear evidence of spatially localized translation. 18 Here we consider only the temporal component of the putative "window."…”

Section: Introductionmentioning

confidence: 99%

Kinetic partitioning during de novo septin filament assembly creates a critical G1 “window of opportunity” for mutant septin function

et al. 2016

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Septin proteins form highly conserved cytoskeletal filaments composed of hetero-oligomers with strict subunit stoichiometry. Mutations within one hetero-oligomerization interface (the "G" interface) bias the mutant septin toward conformations that are incompatible with filament assembly, causing disease in humans and, in budding yeast cells, temperature-sensitive defects in cytokinesis. We previously found that, when the amount of other hetero-oligomerization partners is limiting, wild-type and G interfacemutant alleles of a given yeast septin "compete" along parallel but distinct folding pathways for occupancy of a limited number of positions within septin hetero-octamers. Here, we synthesize a mathematical model that outlines the requirements for this phenomenon: if a wild-type septin traverses a folding pathway that includes a single rate-limiting folding step, the acquisition by a mutant septin of additional slow folding steps creates an initially large disparity between wild-type and mutant in the cellular concentrations of oligomerization-competent monomers. When the 2 alleles are co-expressed, this kinetic disparity results in mutant exclusion from hetero-oligomers, even when the folded mutant monomer is oligomerization-competent. To test this model experimentally, we first visualize the kinetic delay in mutant oligomerization in living cells, and then narrow or widen the "window of opportunity" for mutant septin oligomerization by altering the length of the G1 phase of the yeast cell cycle, and observe the predicted exacerbation or suppression, respectively, of mutant cellular phenotypes. These findings reveal a fundamental kinetic principle governing in vivo assembly of multiprotein complexes, independent of the ability of the subunits to associate with each other.

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“…RIP-chip refers to a strategy in which RNAs in the immunoprecipitate are identified by microarray analysis. The procedure for fission yeast (Amorim et al 2010;Duncan and Mata 2011;Hasan et al 2014) is similar overall to the one used to characterize nuclear ribonucleoprotein particles from Drosophila (see Protocol: Large-Scale Immunopurification of Ribonucleoprotein Complexes from Drosophila Nucleoplasmic Extracts for Tiling Microarrays [Rio 2014c]), mainly differing in the use of whole cell rather than fractionated lysates. High-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) or its variations are now commonly used in other organisms (Darnell 2010), and a CRAC (cross-linking and analysis of cDNA) protocol (similar to HITS-CLIP) used in budding yeast has been adapted by Kilchert et al (2015) to investigate fission yeast RNAs bound to the Mmi1 protein, which targets them for destruction by the exosome.…”

Section: Analysis Of Rna-protein Interactionsmentioning

confidence: 99%