Combined actions of bacteriophage-encoded genes in Wolbachia-induced male lethality

Hiroshi Arai; Hisashi Anbutsu; Yohei Nishikawa; Masato Kogawa; Kazuo Ishii; Masahito Hosokawa; Shiou-Ruei Lin; Masatoshi Ueda; Madoka Nakai; Yasuhisa Kunimi; Toshiyuki Harumoto; Daisuke Kageyama; Haruko Takeyama; Maki N Inoue

doi:10.1016/j.isci.2023.106842

Combined actions of bacteriophage-encoded genes in Wolbachia-induced male lethality

iScience. 2023 May 10;26(6):106842. doi: 10.1016/j.isci.2023.106842. eCollection 2023 Jun 16.

Authors

Hiroshi Arai^{1

2}, Hisashi Anbutsu^{3

4

5}, Yohei Nishikawa^{3

5}, Masato Kogawa⁵, Kazuo Ishii¹, Masahito Hosokawa^{3

5

6

7}, Shiou-Ruei Lin⁸, Masatoshi Ueda¹, Madoka Nakai¹, Yasuhisa Kunimi¹, Toshiyuki Harumoto^{9

10}, Daisuke Kageyama², Haruko Takeyama^{3

5

6

7}, Maki N Inoue¹

Affiliations

¹ United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo 183-8509, Japan.
² National Agriculture and Food Research Organization (NARO), 1-2 Owashi, Tsukuba, Ibaraki 305-0851, Japan.
³ Computational Bio Big-Data Open Innovation Laboratory (CBBD-OIL), National Institute of Advanced Industrial Science and Technology (AIST), 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
⁴ Bioproduction Research Institute, AIST, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
⁵ Research Organization for Nano and Life Innovation, Waseda University, 513 Wasedatsurumaki-cho, Shinjuku-ku, Tokyo 162-0041, Japan.
⁶ Institute for Advanced Research of Biosystem Dynamics, Waseda Research Institute for Science and Engineering, Graduate School of Advanced Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
⁷ Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsucho, Shinjuku-ku, Tokyo 162-8480, Japan.
⁸ Tea Research and Extension Station, 326011 Chung-Hsing RD, Yangmei, Taoyuan, Taiwan, R.O.C.
⁹ Hakubi Center for Advanced Research, Kyoto University. Yoshida-honmachi, Sakyo-ku, Kyoto 606-8501, Japan.
¹⁰ Graduate School of Biostudies, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.

Abstract

Some Wolbachia endosymbionts induce male killing, whereby male offspring of infected females are killed during development; however, the origin and diversity of the underlying mechanisms remain unclear. In this study, we identified a 76 kbp prophage region specific to male-killing Wolbachia hosted by the moth Homona magnanima. The prophage encoded a homolog of the male-killing gene oscar in Ostrinia moths and the wmk gene that induces various toxicities in Drosophila melanogaster. Upon overexpressing these genes in D. melanogaster, wmk-1 and wmk-3 killed all males and most females, whereas Hm-oscar, wmk-2, and wmk-4 had no impact on insect survival. Strikingly, co-expression of tandemly arrayed wmk-3 and wmk-4 killed 90% of males and restored 70% of females, suggesting their conjugated functions for male-specific lethality. While the male-killing gene in the native host remains unknown, our findings highlight the role of bacteriophages in male-killing evolution and differences in male-killing mechanisms among insects.

Keywords: Evolutionary biology; Genetics; Microbiology; Virology.