Defining the Specificity of Cotranslationally Acting Chaperones by Systematic Analysis of mRNAs Associated with Ribosome-Nascent Chain Complexes

Polypeptides exiting the ribosome must fold and assemble in the crowded environment of the cell. Chaperones and other protein homeostasis factors interact with newly translated polypeptides to facilitate their folding and correct localization. Despite the extensive efforts, little is known about the specificity of the chaperones and other factors that bind nascent polypeptides. To address this question we present an approach that systematically identifies cotranslational chaperone substrates through the mRNAs associated with ribosome-nascent chain-chaperone complexes. We here focused on two Saccharomyces cerevisiae chaperones: the Signal Recognition Particle (SRP), which acts cotranslationally to target proteins to the ER, and the Nascent chain Associated Complex (NAC), whose function has been elusive. Our results provide new insights into SRP selectivity and reveal that NAC is a general cotranslational chaperone. We found surprising differential substrate specificity for the three subunits of NAC, which appear to recognize distinct features within nascent chains. Our results also revealed a partial overlap between the sets of nascent polypeptides that interact with NAC and SRP, respectively, and showed that NAC modulates SRP specificity and fidelity in vivo. These findings give us new insight into the dynamic interplay of chaperones acting on nascent chains. The strategy we used should be generally applicable to mapping the specificity, interplay, and dynamics of the cotranslational protein homeostasis network

Monitoring metrical stress in polysyllabic words

Contains fulltext : 55638.pdf (publisher's version ) (Closed access)This study investigated the monitoring of metrical stress information in internally generated speech. In Experiment 1, Dutch participants were asked to judge whether bisyllabic picture names had initial or final stress. Results showed significantly faster decision times for initially stressed targets (e.g., KAno "canoe") than for targets with final stress (e.g., kaNON "cannon"; capital letters indicate stressed syllables). It was demonstrated that monitoring latencies are not a function of the picture naming or object recognition latencies to the same pictures. Experiments 2 and 3 replicated the outcome of the first experiment with trisyllabic picture names. These results are similar to the findings of Wheeldon and Levelt (1995) in a segment monitoring task. The outcome might be interpreted to demonstrate that phonological encoding in speech production is a rightward incremental process. Alternatively, the data might reflect the sequential nature of a perceptual mechanism used to monitor lexical stress.29 p