Structural characterization of a soil viral auxiliary metabolic gene product – a functional chitosanase

Ruonan Wu, Clyde A. Smith, Garry W. Buchko, Ian K. Blaby, David Paez-Espino, Nikos C. Kyrpides, Yasuo Yoshikuni, Jason E. McDermott, Kirsten S. Hofmockel, John R. Cort and Janet K. Jansson ()
Additional contact information
Ruonan Wu: Pacific Northwest National Laboratory
Clyde A. Smith: Stanford University
Garry W. Buchko: Pacific Northwest National Laboratory
Ian K. Blaby: Lawrence Berkeley National Laboratory
David Paez-Espino: Mammoth Biosciences
Nikos C. Kyrpides: Lawrence Berkeley National Laboratory
Yasuo Yoshikuni: Lawrence Berkeley National Laboratory
Jason E. McDermott: Pacific Northwest National Laboratory
Kirsten S. Hofmockel: Pacific Northwest National Laboratory
John R. Cort: Pacific Northwest National Laboratory
Janet K. Jansson: Pacific Northwest National Laboratory

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Metagenomics is unearthing the previously hidden world of soil viruses. Many soil viral sequences in metagenomes contain putative auxiliary metabolic genes (AMGs) that are not associated with viral replication. Here, we establish that AMGs on soil viruses actually produce functional, active proteins. We focus on AMGs that potentially encode chitosanase enzymes that metabolize chitin – a common carbon polymer. We express and functionally screen several chitosanase genes identified from environmental metagenomes. One expressed protein showing endo-chitosanase activity (V-Csn) is crystalized and structurally characterized at ultra-high resolution, thus representing the structure of a soil viral AMG product. This structure provides details about the active site, and together with structure models determined using AlphaFold, facilitates understanding of substrate specificity and enzyme mechanism. Our findings support the hypothesis that soil viruses contribute auxiliary functions to their hosts.

Date: 2022
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DOI: 10.1038/s41467-022-32993-8

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