학술논문
Chicken FMRP Translational Regulator 1 (FMR1) Promotes Early Avian Influenza Virus Transcription without Affecting Viral Progeny Production in DF1 Cells
Document Type
Academic Journal
Author
Source
한국가금학회지. 2021-06 48(2):81-90
Subject
Language
Korean
ISSN
1225-6625
2287-5387
2287-5387
Abstract
조류 인플루엔자 바이러스는 다양한 숙주 단백질을 이용해야만 증식이 가능하다. 포유류 (사람, 쥐) Fragile X mental retardation protein (FMRP)는 최근 인플루엔자 A 바이러스 viral RNP (vRNP)의 조립을 돕고, 이를 핵에서 세포질로 운반시켜 바이러스 증식에 도움을 준다. 하지만, 조류 인플루엔자 바이러스의 주요 숙주인 닭에서는 FMRP translational regulator 1 (FMR1) 유전자의 기능이 규명되지 않았다. 본 연구는 CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/Cas9) 유전자 가위를 이용해 정확히 닭 FMR1 유전자를 제거하여 닭 FMR1 유전자가 조류 인플루엔자 바이러스 증식에 어떤 영향을 끼치는지 연구하였다. 닭 FMR1 유전자는 닭 배아섬유아세포 (DF1세포)에서 초기 조류 인플루엔자 바이러스의 유전자발현을 촉진하나, 감염 후 24시간 뒤에는 바이러스 생산 및 바이러스 중합효소 (Polymerase)의 활성에 영향을 끼치지 않았다. 또한, 야생형 닭 FMR1 유전자를 과발현 함에도 불구하고, 조류 인플루엔자 바이러스의 생산량에는 변화가 없었다. 위 결과들은 닭 FMR1은 포유류 FMR1 유전자에 비해 조류 인플루엔자 바이러스의 증식에 큰 영향을 주지 못하는 숙주인자임을 시사한다. 또한, 닭 FMR1처럼 기존에 포유류에서 알려진 숙주 인자를 목표로 하는 조류 인플루엔자 바이러스 저항성 치료제 및 형질전환 동물을 생산할 때, 조류 시스템에서 위 숙주 인자의 기능이 보존돼 있는지 고찰할 필요가 있다고 사료된다.
Avian influenza viruses (AIVs) must utilize host cellular factors to complete their life cycle, and fragile X mental retardation protein (FMRP) has been reported to be a host factor promoting AIV ribonucleoprotein (vRNP) assembly and exports vRNP from the nucleus to the cytoplasm. The functional role of chicken FMRP translational regulator 1 (cFMR1) as a host factor of AIV is, however, poorly understood. In this study, we targeted the cFMR1 gene in DF1 cells using clustered regularly interspaced short palindromic repeats/Cas9-mediated genome editing to examine the functional role of cFMR1 as a host factor of AIV. We found that cFMR1 stimulated viral gene transcription during early stages of the viruses’ life cycle and did not affect viral progeny production and viral polymerase activity in DF1 cells 24 hours post infection. cFMR1 overexpression did not exert significant effects on virus production, compared to the control. Therefore, unlike in mammalian systems (e.g., humans or mice), cFMR1 did not play a pivotal role in AIV but only seemed to stimulate viral proliferation during early stages of the viral life cycle. These results imply that the interplay between host factors and AIV differs between mammals and avian species, and such differences should be considered when developing anti-viral drugs for birds or establishing AIV-resistant bird models.
Avian influenza viruses (AIVs) must utilize host cellular factors to complete their life cycle, and fragile X mental retardation protein (FMRP) has been reported to be a host factor promoting AIV ribonucleoprotein (vRNP) assembly and exports vRNP from the nucleus to the cytoplasm. The functional role of chicken FMRP translational regulator 1 (cFMR1) as a host factor of AIV is, however, poorly understood. In this study, we targeted the cFMR1 gene in DF1 cells using clustered regularly interspaced short palindromic repeats/Cas9-mediated genome editing to examine the functional role of cFMR1 as a host factor of AIV. We found that cFMR1 stimulated viral gene transcription during early stages of the viruses’ life cycle and did not affect viral progeny production and viral polymerase activity in DF1 cells 24 hours post infection. cFMR1 overexpression did not exert significant effects on virus production, compared to the control. Therefore, unlike in mammalian systems (e.g., humans or mice), cFMR1 did not play a pivotal role in AIV but only seemed to stimulate viral proliferation during early stages of the viral life cycle. These results imply that the interplay between host factors and AIV differs between mammals and avian species, and such differences should be considered when developing anti-viral drugs for birds or establishing AIV-resistant bird models.