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Mezhyrova et al. Microbiome Res Rep 2023;2:28 Microbiome Research
DOI: 10.20517/mrr.2023.28
Reports

Original Article Open Access

In vitro characterization of the phage lysis protein
MS2-L

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Julija Mezhyrova , Janosch Martin , Clara Börnsen , Volker Dötsch , Achilleas Stefanos Frangakis , Nina
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Morgner , Frank Bernhard 1
1
Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Goethe University, Frankfurt am Main
60438, Germany.
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Institute of Physical and Theoretical Chemistry, Goethe University, Frankfurt am Main 60438, Germany.
3
Buchmann Institute for Molecular Life Sciences & Institute of Biophysics, Goethe University, Frankfurt am Main 60438,
Germany.
Correspondence to: Dr. Frank Bernhard, Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance,
Goethe University, Max von Laue Str. 9, Frankfurt am Main 60438, Germany. E-mail: fbern@bpc.uni-frankfurt.de
How to cite this article: Mezhyrova J, Martin J, Börnsen C, Dötsch V, Frangakis AS, Morgner N, Bernhard F. In vitro
characterization of the phage lysis protein MS2-L. Microbiome Res Rep 2023;2:28. https://dx.doi.org/10.20517/mrr.2023.28
Received: 19 Apr 2023 First Decision: 25 Jun 2023 Revised: 29 Jun 2023 Accepted: 10 Jul 2023 Published: 20 Jul 2023

Academic Editor: Douwe van Sinderen Copy Editor: Dong-Li Li Production Editor: Dong-Li Li

Abstract
Background: The peptide MS2-L represents toxins of the ssRNA Leviviridae phage family and consists of a
predicted N-terminal soluble domain followed by a transmembrane domain. MS2-L mediates bacterial cell lysis
through the formation of large lesions in the cell envelope, but further details of this mechanism as a prerequisite
for applied bioengineering studies are lacking. The chaperone DnaJ is proposed to modulate MS2-L activity,
whereas other cellular targets of MS2-L are unknown.

Methods: Here, we provide a combined in vitro and in vivo overexpression approach to reveal molecular insights
into MS2-L action and its interaction with DnaJ. Full-length MS2-L and truncated derivatives were synthesized cell-
free and co-translationally inserted into nanodiscs or solubilized in detergent micelles. By native liquid bead ion
desorption mass spectrometry, we demonstrate that MS2-L assembles into high oligomeric states after membrane
insertion.

Results: Oligomerization is directed by the transmembrane domain and is impaired in detergent environments.
Studies with truncated MS2-L derivatives provide evidence that the soluble domain acts as a modulator of oligomer
formation. DnaJ strongly interacts with MS2-L in membranes as well as in detergent environments. However, this
interaction affects neither the MS2-L membrane insertion efficiency nor its oligomerization in nanodisc

© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0
International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing,
adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as
long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and
indicate if changes were made.

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