Asp149 and Asp152 in chicken and human ANP32A play an essential role in the interaction with influenza viral polymerase.

Park, Young Hyun; Woo, Seung Je; Chungu, Kelly; Lee, Su Bin; Shim, Ji Hyeon; Lee, Hong Jo; Kim, Iktae; Rengaraj, Deivendran; Song, Chang-Seon; Suh, Jeong-Yong; Lim, Jeong Mook; Han, Jae Yong

Park, Young Hyun; Woo, Seung Je; Chungu, Kelly; Lee, Su Bin; Shim, Ji Hyeon; Lee, Hong Jo; Kim, Iktae; Rengaraj, Deivendran; Song, Chang-Seon; Suh, Jeong-Yong; Lim, Jeong Mook; Han, Jae Yong.

Afiliación

Park YH; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Woo SJ; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Chungu K; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Lee SB; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Shim JH; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Lee HJ; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Kim I; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Rengaraj D; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Song CS; Avian Diseases Laboratory, College of Veterinary Medicine, Konkuk University, Seoul, Republic of Korea.
Suh JY; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Lim JM; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Han JY; Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.

FASEB J ; 35(6): e21630, 2021 06.

Article en En | MEDLINE | ID: mdl-33982347

RESUMEN

The acidic nuclear phosphoprotein 32 family member A (ANP32A) is a cellular host factor that determines the host tropism of the viral polymerase (vPol) of avian influenza viruses (AIVs). Compared with human ANP32A (hANP32A), chicken ANP32A contains an additional 33 amino acid residues (176-208) duplicated from amino acid residues 149-175 (27 residues), suggesting that these residues could be involved in increasing vPol activity by strengthening interactions between ANP32A and vPol. However, the molecular interactions and functional roles of the 27 residues within hANP32A during AIV vPol activity remain unclear. Here, we examined the functional role of 27 residues of hANP32A based on comparisons with other human (h) ANP32 family members. It was notable that unlike hANP32A and hANP32B, hANP32C could not support vPol activity or replication of AIVs, despite the fact that hANP32C shares a higher sequence identity with hANP32A than hANP32B. Pairwise comparison between hANP32A and hANP32C revealed that Asp149 (D149) and Asp152 (D152) are involved in hydrogen bonding and electrostatic interactions, respectively, which support vPol activity. Mutation of these residues reduced the interaction between hANP32A and vPol. Finally, we demonstrated that precise substitution of the identified residues within chicken ANP32A via homology-directed repair using the CRISPR/Cas9 system resulted in a marked reduction of viral replication in chicken cells. These results increase our understanding of ANP32A function and may facilitate the development of AIV-resistant chickens via precise modification of residues within ANP32A.

Asunto(s)

Ácido Aspártico/metabolismo; ADN Polimerasa Dirigida por ADN/metabolismo; Virus de la Influenza A/enzimología; Mutación; Proteínas Nucleares/metabolismo; Infecciones por Orthomyxoviridae/virología; Proteínas de Unión al ARN/metabolismo; Proteínas Virales/metabolismo; Secuencia de Aminoácidos; Animales; Ácido Aspártico/química; Ácido Aspártico/genética; Pollos; ADN Polimerasa Dirigida por ADN/genética; Humanos; Proteínas Nucleares/química; Proteínas Nucleares/genética; Infecciones por Orthomyxoviridae/metabolismo; Proteínas de Unión al ARN/química; Proteínas de Unión al ARN/genética; Homología de Secuencia; Proteínas Virales/genética

Palabras clave

ANP32A; host factor; influenza virus; precise genome editing; protein-protein interaction

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Virus de la Influenza A / Proteínas Virales / Proteínas Nucleares / Proteínas de Unión al ARN / Ácido Aspártico / Infecciones por Orthomyxoviridae / ADN Polimerasa Dirigida por ADN / Mutación Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: FASEB J Asunto de la revista: BIOLOGIA / FISIOLOGIA Año: 2021 Tipo del documento: Article Pais de publicación: Estados Unidos

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