Mouse APOBEC3 Expression in NIH 3T3 Cells Mediates Hypermutation of AKV Murine Leukemia Virus

Overview

Journal Virology

Specialty Microbiology

Date 2018 Apr 2

PMID 29605684

Citations 5

Authors

Stefano Boi

Morgan E Ferrell

Ming Zhao

Kim J Hasenkrug

Leonard H Evans

Affiliations

Soon will be listed here.

Abstract

Mouse APOBEC3 (mA3) is a cytidine deaminase that can act on the single-stranded DNA reverse transcripts of retroviruses resulting in G→A hypermutation of proviral DNA. Many mA3 studies have used NIH 3T3 cells assuming that endogenous mA3 production was negligible. We developed a monoclonal antibody specific for mA3 that reveals detectable mA3 in NIH 3T3 cells and we demonstrate that AKV released from the cells undergoes G→A hypermutation. Inactivation of the mA3 gene abolished the deamination confirming that AKV hypermutation was mediated by mA3. The G→A mutations in AKV viral transcripts deviated from a normal distribution with all the mutations contained within only 20% of the transcripts. Single cell analyses revealed that the expression of mA3 in NIH 3T3 cells was limited to 20% of the cells, which likely accounted for the abnormal distribution of mutations. Endogenous NIH 3T3 mA3 was found to restrict AKV replication.

Citing Articles

Apobec-mediated retroviral hypermutation in vivo is dependent on mouse strain.

Byun H, Singh G, Xu W, Das P, Reyes A, Battenhouse A PLoS Pathog. 2024; 20(8):e1012505.

PMID: 39208378 PMC: 11389910. DOI: 10.1371/journal.ppat.1012505.

The Role of APOBECs in Viral Replication.

Xu W, Byun H, Dudley J Microorganisms. 2020; 8(12).

PMID: 33266042 PMC: 7760323. DOI: 10.3390/microorganisms8121899.

Mouse APOBEC3 Restriction of Retroviruses.

Salas-Briceno K, Zhao W, Ross S Viruses. 2020; 12(11).

PMID: 33121095 PMC: 7692085. DOI: 10.3390/v12111217.

A Protein Antagonist of Activation-Induced Cytidine Deaminase Encoded by a Complex Mouse Retrovirus.

Singh G, Byun H, Ali A, Medina F, Wylie D, Shivram H mBio. 2019; 10(4).

PMID: 31409681 PMC: 6692512. DOI: 10.1128/mBio.01678-19.

Friend retrovirus studies reveal complex interactions between intrinsic, innate and adaptive immunity.

Dittmer U, Sutter K, Kassiotis G, Zelinskyy G, Banki Z, Stoiber H FEMS Microbiol Rev. 2019; 43(5):435-456.

PMID: 31087035 PMC: 6735856. DOI: 10.1093/femsre/fuz012.

References

Portis J, Spangrude G, McAtee F . Identification of a sequence in the unique 5' open reading frame of the gene encoding glycosylated Gag which influences the incubation period of neurodegenerative disease induced by a murine retrovirus. J Virol. 1994; 68(6):3879-87. PMC: 236893. DOI: 10.1128/JVI.68.6.3879-3887.1994. View

Stavrou S, Nitta T, Kotla S, Ha D, Nagashima K, Rein A . Murine leukemia virus glycosylated Gag blocks apolipoprotein B editing complex 3 and cytosolic sensor access to the reverse transcription complex. Proc Natl Acad Sci U S A. 2013; 110(22):9078-83. PMC: 3670389. DOI: 10.1073/pnas.1217399110. View

Gartner K, Wiktorowicz T, Park J, Mergia A, Rethwilm A, Scheller C . Accuracy estimation of foamy virus genome copying. Retrovirology. 2009; 6:32. PMC: 2678077. DOI: 10.1186/1742-4690-6-32. View

Browne E, Littman D . Species-specific restriction of apobec3-mediated hypermutation. J Virol. 2007; 82(3):1305-13. PMC: 2224457. DOI: 10.1128/JVI.01371-07. View

Gerpe M, Renner T, Belanger K, Lam C, Aydin H, Langlois M . N-linked glycosylation protects gammaretroviruses against deamination by APOBEC3 proteins. J Virol. 2014; 89(4):2342-57. PMC: 4338886. DOI: 10.1128/JVI.03330-14. View

Bishop K, Holmes R, Sheehy A, Davidson N, Cho S, Malim M . Cytidine deamination of retroviral DNA by diverse APOBEC proteins. Curr Biol. 2004; 14(15):1392-6. DOI: 10.1016/j.cub.2004.06.057. View

Chiu Y, Greene W . The APOBEC3 cytidine deaminases: an innate defensive network opposing exogenous retroviruses and endogenous retroelements. Annu Rev Immunol. 2008; 26:317-53. DOI: 10.1146/annurev.immunol.26.021607.090350. View

Doehle B, Schafer A, Wiegand H, Bogerd H, Cullen B . Differential sensitivity of murine leukemia virus to APOBEC3-mediated inhibition is governed by virion exclusion. J Virol. 2005; 79(13):8201-7. PMC: 1143768. DOI: 10.1128/JVI.79.13.8201-8207.2005. View

Boi S, Kolokithas A, Shepard J, Linwood R, Rosenke K, Van Dis E . Incorporation of mouse APOBEC3 into murine leukemia virus virions decreases the activity and fidelity of reverse transcriptase. J Virol. 2014; 88(13):7659-62. PMC: 4054435. DOI: 10.1128/JVI.00967-14. View

10.

Xu H, Chertova E, Chen J, Ott D, Roser J, Hu W . Stoichiometry of the antiviral protein APOBEC3G in HIV-1 virions. Virology. 2006; 360(2):247-56. DOI: 10.1016/j.virol.2006.10.036. View

11.

Pathak V, Temin H . Broad spectrum of in vivo forward mutations, hypermutations, and mutational hotspots in a retroviral shuttle vector after a single replication cycle: substitutions, frameshifts, and hypermutations. Proc Natl Acad Sci U S A. 1990; 87(16):6019-23. PMC: 54463. DOI: 10.1073/pnas.87.16.6019. View

12.

Langlois M, Kemmerich K, Rada C, Neuberger M . The AKV murine leukemia virus is restricted and hypermutated by mouse APOBEC3. J Virol. 2009; 83(22):11550-9. PMC: 2772689. DOI: 10.1128/JVI.01430-09. View

13.

Schneider C, Rasband W, Eliceiri K . NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012; 9(7):671-5. PMC: 5554542. DOI: 10.1038/nmeth.2089. View

14.

Esnault C, Millet J, Schwartz O, Heidmann T . Dual inhibitory effects of APOBEC family proteins on retrotransposition of mammalian endogenous retroviruses. Nucleic Acids Res. 2006; 34(5):1522-31. PMC: 1401513. DOI: 10.1093/nar/gkl054. View

15.

Abudu A, Takaori-Kondo A, Izumi T, Shirakawa K, Kobayashi M, Sasada A . Murine retrovirus escapes from murine APOBEC3 via two distinct novel mechanisms. Curr Biol. 2006; 16(15):1565-70. DOI: 10.1016/j.cub.2006.06.055. View

16.

Rulli Jr S, Mirro J, Hill S, Lloyd P, Gorelick R, Coffin J . Interactions of murine APOBEC3 and human APOBEC3G with murine leukemia viruses. J Virol. 2008; 82(13):6566-75. PMC: 2447093. DOI: 10.1128/JVI.01357-07. View

17.

Rowe W, Hartley J, Lander M, PUGH W, Teich N . Noninfectious AKR mouse embryo cell lines in which each cell has the capacity to be activated to produce infectious murine leukemia virus. Virology. 1971; 46(3):866-76. DOI: 10.1016/0042-6822(71)90087-0. View

18.

Chesebro B, Britt W, Evans L, Wehrly K, Nishio J, CLOYD M . Characterization of monoclonal antibodies reactive with murine leukemia viruses: use in analysis of strains of friend MCF and Friend ecotropic murine leukemia virus. Virology. 1983; 127(1):134-48. DOI: 10.1016/0042-6822(83)90378-1. View

19.

Paprotka T, Venkatachari N, Chaipan C, Burdick R, Delviks-Frankenberry K, Hu W . Inhibition of xenotropic murine leukemia virus-related virus by APOBEC3 proteins and antiviral drugs. J Virol. 2010; 84(11):5719-29. PMC: 2876585. DOI: 10.1128/JVI.00134-10. View

20.

Sova P, Volsky D . Efficiency of viral DNA synthesis during infection of permissive and nonpermissive cells with vif-negative human immunodeficiency virus type 1. J Virol. 1993; 67(10):6322-6. PMC: 238061. DOI: 10.1128/JVI.67.10.6322-6326.1993. View