Peroxin Pex21p Interacts with C-terminal Noncatalytic Domain of Yeast Seryl-tRNA Synthetase and Forms a Specific Ternary Complex with tRNASer

Document Type

Article

Publication Date

4-20-2007

Abstract

The seryl‐tRNA synthetase from Saccharomyces cerevisiae interacts with the peroxisome biogenesis‐related factor Pex21p. Several deletion mutants of seryl‐tRNA synthetase were constructed and inspected for their ability to interact with Pex21p in a yeast two‐hybrid assay, allowing mapping of the synthetase domain required for complex assembly. Deletion of the 13 C‐terminal amino acids abolished Pex21p binding to seryl‐tRNA synthetase. The catalytic parameters of purified truncated seryl‐tRNA synthetase, determined in the serylation reaction, were found to be almost identical to those of the native enzyme. In vivo loss of interaction with Pex21p was confirmed in vitro by coaffinity purification. These data indicate that the C‐terminally appended domain of yeast seryl‐tRNA synthetase does not participate in substrate binding, but instead is required for association with Pex21p. We further determined that Pex21p does not directly bind tRNA, and nor does it possess a tRNA‐binding motif, but it instead participates in the formation of a specific ternary complex with seryl‐tRNA synthetase and tRNASer, strengthening the interaction of seryl‐tRNA synthetase with its cognate tRNASer.

Comments

This article was originally published in FEBS Journal, volume 274, in 2007. https://doi.org/10.1111/j.1742-4658.2007.05812.x

Copyright

Wiley

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