Determination of the factors responsible for the tropism of SARS-CoV-2-related bat coronaviruses to
Rhinolophus
bat ACE2
Authors: Fujita Shigeru , Kosugi Yusuke , Kimura Izumi , Tokunaga Kenzo ,   The Genotype to Phenotype Japan (G2P-Japan) Consortium, Ito Jumpei , Sato Kei , Matsuno Keita , Nao Naganori , Sawa Hirofumi , Tanaka Shinya , Tsuda Masumi , Wang Lei , Oda Yoshikata , Ferdous Zannatul , Shishido Kenji , Fukuhara Takasuke , Tamura Tomokazu , Suzuki Rigel , Suzuki Saori , Ito Hayato , Kaku Yu , Misawa Naoko , Plianchaisuk Arnon , Guo Ziyi , Hinay Alfredo A. , Uriu Keiya , Tolentino Jarel Elgin M. , Chen Luo , Pan Lin , Suganami Mai , Chiba Mika , Yoshimura Ryo , Yasuda Kyoko , Iida Keiko , Ohsumi Naomi , Strange Adam P. , Tanaka Shiho , Yoshimura Kazuhisa , Sadamasu Kenji , Nagashima Mami , Asakura Hiroyuki , Yoshida Isao , Nakagawa So , Takaori-Kondo Akifumi , Nagata Kayoko , Nomura Ryosuke , Horisawa Yoshihito , Tashiro Yusuke , Kawai Yugo , Takayama Kazuo , Hashimoto Rina , Deguchi Sayaka , Watanabe Yukio , Sakamoto Ayaka , Yasuhara Naoko , Hashiguchi Takao , Suzuki Tateki , Kimura Kanako , Sasaki Jiei , Nakajima Yukari , Yajima Hisano , Irie Takashi , Kawabata Ryoko , Tabata Kaori , Ikeda Terumasa , Nasser Hesham , Shimizu Ryo , Monira Begum M. S. T. , Jonathan Michael , Mugita Yuka , Takahashi Otowa , Ichihara Kimiko , Motozono Chihiro , Ueno Takamasa , Toyoda Mako , Saito Akatsuki , Shofa Maya , Shibatani Yuki , Nishiuchi Tomoko , Shirakawa Kotaro 
Published: September 2023

Differences in host angiotensin converting enzyme 2 (ACE2) genes may affect the host range of SARS-CoV-2-related coronaviruses (SC2r-CoVs) and further determine the tropism of host ACE2 for the infection receptor. However, the factor(s) responsible for determining the host tropism of SC2r-CoVs, which may in part be determined by the tropism of host ACE2 usage, remains unclear. Here, we use the pseudoviruses with the spike proteins of two Laotian SC2r-CoVs, BANAL-20-236 and BANAL-20-52, and the cells expressing ACE2 proteins of eight different
Rhinolophus
bat species to show that these two spikes have different tropisms for
Rhinolophus
bat ACE2. Through structural analysis and cell culture experiments, we demonstrate that this tropism is determined by residue 493 of the spike and residues 31 and 35 of ACE2. Our results suggest that SC2r-CoVs exhibit differential ACE2 tropism, which may be driven by adaptation to different
Rhinolophus
bat ACE2 proteins.

IMPORTANCE

The efficiency of infection receptor use is the first step in determining the species tropism of viruses. After the coronavirus disease 2019 pandemic, a number of SARS-CoV-2-related coronaviruses (SC2r-CoVs) were identified in
Rhinolophus
bats, and some of them can use human angiotensin converting enzyme 2 (ACE2) for the infection receptor without acquiring additional mutations. This means that the potential of certain SC2r-CoVs to cause spillover from bats to humans is "off-the-shelf." However, both SC2r-CoVs and
Rhinolophus
bat species are highly diversified, and the host tropism of SC2r-CoVs remains unclear. Here, we focus on two Laotian SC2r-CoVs, BANAL-20-236 and BANAL-20-52, and determine how the tropism of SC2r-CoVs to
Rhinolophus
bat ACE2 is determined at the amino acid resolution level.