Nonspecific N-terminal tetrapeptide insertions disrupt the translation arrest induced by ribosome-arresting peptide sequences

Akinao Kobo; Hideki Taguchi; Yuhei Chadani

doi:10.1016/j.jbc.2024.107360

Nonspecific N-terminal tetrapeptide insertions disrupt the translation arrest induced by ribosome-arresting peptide sequences

J Biol Chem. 2024 May 11;300(6):107360. doi: 10.1016/j.jbc.2024.107360. Online ahead of print.

Authors

Akinao Kobo¹, Hideki Taguchi², Yuhei Chadani³

Affiliations

¹ School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
² School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan; Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan. Electronic address: taguchi@bio.titech.ac.jp.
³ Faculty of Environmental, Life, Natural Science and Technology, Okayama University, Okayama, Japan. Electronic address: ychadani@okayama-u.ac.jp.

PMID: 38735477
DOI: 10.1016/j.jbc.2024.107360

Abstract

The nascent polypeptide chains passing through the ribosome tunnel not only serve as an intermediate of protein synthesis but also, in some cases, act as dynamic genetic information, controlling translation through interaction with the ribosome. One notable example is Escherichia coli SecM, in which translation of the ribosome arresting peptide (RAP) sequence in SecM leads to robust elongation arrest. Translation regulations, including the SecM-induced translation arrest, play regulatory roles such as gene expression control. Recent investigations have indicated that the insertion of a peptide sequence, SKIK (or MSKIK), into the adjacent N-terminus of the RAP sequence of SecM behaves as an "arrest canceler". As the study did not provide a direct assessment of the strength of translation arrest, we conducted detailed biochemical analyses. The results revealed that the effect of SKIK insertion on weakening SecM-induced translation arrest was not specific to the SKIK sequence, that is, other tetrapeptide sequences inserted just before the RAP sequence also attenuated the arrest. Our data suggest that SKIK or other tetrapeptide insertions disrupt the context of the RAP sequence rather than canceling or preventing the translation arrest.

Keywords: CmlA leader; SKIK tag; SecM; difficult-to-express proteins; nascent chain; ribosomal stalling; ribosome arrest peptide; translation arrest; translation regulation.