AUTHOR=Lee Junbeom , Cha Wook Hyun , Lee Dae-Weon 

TITLE=Multiple Precursor Proteins of Thanatin Isoforms, an Antimicrobial Peptide Associated With the Gut Symbiont of Riptortus pedestris

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.796548

DOI=10.3389/fmicb.2021.796548

ISSN=1664-302X

ABSTRACT=<p>Thanatin is an antimicrobial peptide (AMP) generated by insects for defense against bacterial infections. In the present study, we performed cDNA cloning of thanatin and found the presence of multiple precursor proteins from the bean bug, <italic>Riptortus pedestris</italic>. The cDNA sequences encoded 38 precursor proteins, generating 13 thanatin isoforms. In the phylogenetic analysis, thanatin isoforms were categorized into two groups based on the presence of the membrane attack complex/perforin (MACPF) domain. In insect-bacterial symbiosis, specific substances are produced by the immune system of the host insect and are known to modulate the symbiont’s population. Therefore, to determine the biological function of thanatin isoforms in symbiosis, the expression levels of three AMP genes were compared between aposymbiotic insects and symbiotic <italic>R. pedestris</italic>. The expression levels of the <italic>thanatin</italic> genes were significantly increased in the M4 crypt, a symbiotic organ, of symbiotic insects upon systemic bacterial injection. Further, synthetic thanatin isoforms exhibited antibacterial activity against gut-colonized <italic>Burkholderia</italic> symbionts rather than <italic>in vitro</italic>-cultured <italic>Burkholderia</italic> cells. Interestingly, the suppression of <italic>thanatin</italic> genes significantly increased the population of <italic>Burkholderia</italic> gut symbionts in the M4 crypt under systemic <italic>Escherichia coli</italic> K12 injection. Overgrown <italic>Burkholderia</italic> gut symbionts were observed in the hemolymph of host insects and exhibited insecticidal activity. Taken together, these results suggest that thanatin of <italic>R. pedestris</italic> is a host-derived symbiotic factor and an AMP that controls the population of gut-colonized <italic>Burkholderia</italic> symbionts.</p>