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Custom Vector Construction> | ...pET28a-based vectors for bacterial expression of His6-tagged human Npl4, and wt, TAQ244-246/TAL and TAQ244-246/GGQ mutant forms of ANKZF1 were prepared by GenScript (Piscataway, NJ). pcDNA3.1+/ C-(K)DYK-based vectors for expression in human cells of C-terminally FLAG tagged human TCF25 and ANKZF1 were made using the CloneEZ strategy (GenScript)... | Get A Quote |
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The ribosome-associated quality control (RQC) pathway degrades nascent chains (NCs) arising from interrupted translation. First, recycling factors split stalled ribosomes, yielding NC-tRNA/60S ribosome-nascent chain complexes (60S RNCs). 60S RNCs associate with NEMF, which recruits the E3 ubiquitin ligase Listerin that ubiquitinates NCs. The mechanism of subsequent ribosomal release of Ub-NCs remains obscure. We found that, in non-ubiquitinated 60S RNCs and 80S RNCs formed on non-stop mRNAs, tRNA is not firmly fixed in the P site, which allows peptidyl-tRNA hydrolase Ptrh1 to cleave NC-tRNA, suggesting the existence of a pathway involving release of non-ubiquitinated NCs. Association with NEMF and Listerin and ... More