Comparative genomics and proteomic analyses between lethal and nonlethal strains of Plasmodium berghei Mamoru Niikura a, 1 , ShineIchi Inoue a , Toshiyuki Fukutomi b , Junya Yamagishi c , Hiroko Asahi a , Fumie Kobayashi a, *, 1 a Department of Infectious Diseases, Kyorin University School of Medicine, Tokyo 181e8611, Japan b Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Mitaka, Tokyo 181-8611, Japan c Division of Collaboration and Education, Research Center for Zoonosis Control, Hokkaido University, North 20, West 10 Kita-ku, Sapporo, Hokkaido 001- 0020, Japan highlights graphical abstract  Plasmodium berghei XAT is deficient in the RAP complex.  sbp1 , which is associated with sequestration, is deleted in Pb XAT.  Pb XAT may not produce mature male and female gametocytes.  RAP2/3 may be involved in virulence of malaria, independently of RAP1. article info Article history: Received 24 July 2017 Received in revised form 16 November 2017 Accepted 2 January 2018 Available online 4 January 2018 Keywords: Plasmodium Comparative genomics Proteomics RAP1 RAP2/3 abstract Plasmodium berghei (Pb) XAT, a rodent malaria parasite, is an irradiation-attenuated variant derived from the lethal strain Pb NK65. Differences in genome sequence, protein structure and function between Pb XAT and Pb NK65 are currently unknown. In this study, to investigate genetic alterations in Pb XAT, we performed comparative genomics and proteomics analyses of nonlethal and lethal strains of Pb. We found mutations, such as a deletion mutation in rhoptry-associated protein (rap) 1 , and deletion of rap2/3 and skeleton-binding protein 1 (sbp1), in Pb XAT. RAP1 is required for targeting of RAP2 to the rhoptries. However, the contribution of RAP2/3 to the lethality of Plasmodium is unclear. Therefore, we generated RAP1- and RAP2/3-deficient mutants of Pb ANKA, a reference strain of P. berghei. Furthermore, we investigated the effect of RAP1 and RAP2/3 deficiency on the outcome of infection. The parasitemia in mice infected with RAP1-deficient parasites was increased compared to that in control parasite-infected mice during the early phase of infection. However, mice infected with RAP1-deficient parasites survived longer than did control parasite-infected mice. Moreover, mice infected with RAP2/3-deficient parasites showed low levels of parasitemia and ultimately recovered from the infection The aim of this study was to investigate the effect of RAP2/3 expression on the outcome of infection with Pb XAT using a RAP2/3- expressing Pb XAT. Results showed that complementation of RAP2/3 expression in Pb XAT partially * Corresponding author. Department of Infectious Diseases, Kyorin University School of Medicine, 6e20e2 Shinkawa, Mitaka, Tokyo 181e8611, Japan. E-mail address: fumfum@ks.kyorin-u.ac.jp (F. Kobayashi). 1 These authors contributed equally to this work. Contents lists available at ScienceDirect Experimental Parasitology journal homepage: www.elsevier.com/locate/yexpr https://doi.org/10.1016/j.exppara.2018.01.001 0014-4894/© 2018 Elsevier Inc. All rights reserved. Experimental Parasitology 185 (2018) 1e9