Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein

Kathy R. Chaurasiya; Micah J. McCauley; Wei Wang; Dominic F. Qualley; Tiyun Wu; Shingo Kitamura; Hylkje Geertsema; Denise S.B. Chan; Amber Hertz; Yasumasa Iwatani; Judith G. Levin; Karin Musier-Forsyth; Ioulia F Rouzina; Mark C. Williams

doi:10.1038/nchem.1795

Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein

Kathy R. Chaurasiya
, Micah J. McCauley
, Wei Wang
, Dominic F. Qualley
, Tiyun Wu
, Shingo Kitamura
, Hylkje Geertsema
, Denise S.B. Chan
, Amber Hertz
, Yasumasa Iwatani
, Judith G. Levin
, Karin Musier-Forsyth
, Ioulia F Rouzina
, Mark C. Williams

Biochemistry, Molecular Biology, and Biophysics (CBS)

Research output: Contribution to journal › Article › peer-review

64 Scopus citations

Abstract

The human APOBEC3 proteins are a family of DNA-editing enzymes that play an important role in the innate immune response against retroviruses and retrotransposons. APOBEC3G is a member of this family that inhibits HIV-1 replication in the absence of the viral infectivity factor Vif. Inhibition of HIV replication occurs by both deamination of viral single-stranded DNA and a deamination-independent mechanism. Efficient deamination requires rapid binding to and dissociation from ssDNA. However, a relatively slow dissociation rate is required for the proposed deaminase-independent roadblock mechanism in which APOBEC3G binds the viral template strand and blocks reverse transcriptase- catalysed DNA elongation. Here, we show that APOBEC3G initially binds ssDNA with rapid on-off rates and subsequently converts to a slowly dissociating mode. In contrast, an oligomerization-deficient APOBEC3G mutant did not exhibit a slow off rate. We propose that catalytically active monomers or dimers slowly oligomerize on the viral genome and inhibit reverse transcription.

Original language	English (US)
Pages (from-to)	28-33
Number of pages	6
Journal	Nature Chemistry
Volume	6
Issue number	1
DOIs	https://doi.org/10.1038/nchem.1795
State	Published - Jan 2014

Bibliographical note

Funding Information:
The authors thank D. Pollpeter, M.H. Malim and D. Rueda for valuable discussions, and M.F. Goodman for his generous gift of the F126A/W127A mutant clone. This work was supported in part by the National Institutes of Health (GM072462 to M.C.W. and GM065056 to K.M-F.) and the National Science Foundation (MCB-1243883 to M.C.W.), the Japan Society for the Promotion of Science (JSPS; KAKENHI_24590568 to Y.I.), and in part by funds from the NIH Intramural Research Program (NICHD; to J.G.L.). K.R.C. was supported by the NSF IGERT Program (DGE-0504331).

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Chaurasiya, K. R., McCauley, M. J., Wang, W., Qualley, D. F., Wu, T., Kitamura, S., Geertsema, H., Chan, D. S. B., Hertz, A., Iwatani, Y., Levin, J. G., Musier-Forsyth, K., Rouzina, I. F., & Williams, M. C. (2014). Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein. Nature Chemistry, 6(1), 28-33. https://doi.org/10.1038/nchem.1795

Chaurasiya, KR, McCauley, MJ, Wang, W, Qualley, DF, Wu, T, Kitamura, S, Geertsema, H, Chan, DSB, Hertz, A, Iwatani, Y, Levin, JG, Musier-Forsyth, K, Rouzina, IF & Williams, MC 2014, 'Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein', Nature Chemistry, vol. 6, no. 1, pp. 28-33. https://doi.org/10.1038/nchem.1795

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title = "Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein",

abstract = "The human APOBEC3 proteins are a family of DNA-editing enzymes that play an important role in the innate immune response against retroviruses and retrotransposons. APOBEC3G is a member of this family that inhibits HIV-1 replication in the absence of the viral infectivity factor Vif. Inhibition of HIV replication occurs by both deamination of viral single-stranded DNA and a deamination-independent mechanism. Efficient deamination requires rapid binding to and dissociation from ssDNA. However, a relatively slow dissociation rate is required for the proposed deaminase-independent roadblock mechanism in which APOBEC3G binds the viral template strand and blocks reverse transcriptase- catalysed DNA elongation. Here, we show that APOBEC3G initially binds ssDNA with rapid on-off rates and subsequently converts to a slowly dissociating mode. In contrast, an oligomerization-deficient APOBEC3G mutant did not exhibit a slow off rate. We propose that catalytically active monomers or dimers slowly oligomerize on the viral genome and inhibit reverse transcription.",

author = "Chaurasiya, \{Kathy R.\} and McCauley, \{Micah J.\} and Wei Wang and Qualley, \{Dominic F.\} and Tiyun Wu and Shingo Kitamura and Hylkje Geertsema and Chan, \{Denise S.B.\} and Amber Hertz and Yasumasa Iwatani and Levin, \{Judith G.\} and Karin Musier-Forsyth and Rouzina, \{Ioulia F\} and Williams, \{Mark C.\}",

note = "Funding Information: The authors thank D. Pollpeter, M.H. Malim and D. Rueda for valuable discussions, and M.F. Goodman for his generous gift of the F126A/W127A mutant clone. This work was supported in part by the National Institutes of Health (GM072462 to M.C.W. and GM065056 to K.M-F.) and the National Science Foundation (MCB-1243883 to M.C.W.), the Japan Society for the Promotion of Science (JSPS; KAKENHI\_24590568 to Y.I.), and in part by funds from the NIH Intramural Research Program (NICHD; to J.G.L.). K.R.C. was supported by the NSF IGERT Program (DGE-0504331).",

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AU - Chaurasiya, Kathy R.

AU - McCauley, Micah J.

AU - Wang, Wei

AU - Qualley, Dominic F.

AU - Wu, Tiyun

AU - Kitamura, Shingo

AU - Geertsema, Hylkje

AU - Chan, Denise S.B.

AU - Hertz, Amber

AU - Iwatani, Yasumasa

AU - Levin, Judith G.

AU - Musier-Forsyth, Karin

AU - Rouzina, Ioulia F

AU - Williams, Mark C.

N1 - Funding Information: The authors thank D. Pollpeter, M.H. Malim and D. Rueda for valuable discussions, and M.F. Goodman for his generous gift of the F126A/W127A mutant clone. This work was supported in part by the National Institutes of Health (GM072462 to M.C.W. and GM065056 to K.M-F.) and the National Science Foundation (MCB-1243883 to M.C.W.), the Japan Society for the Promotion of Science (JSPS; KAKENHI_24590568 to Y.I.), and in part by funds from the NIH Intramural Research Program (NICHD; to J.G.L.). K.R.C. was supported by the NSF IGERT Program (DGE-0504331).

PY - 2014/1

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N2 - The human APOBEC3 proteins are a family of DNA-editing enzymes that play an important role in the innate immune response against retroviruses and retrotransposons. APOBEC3G is a member of this family that inhibits HIV-1 replication in the absence of the viral infectivity factor Vif. Inhibition of HIV replication occurs by both deamination of viral single-stranded DNA and a deamination-independent mechanism. Efficient deamination requires rapid binding to and dissociation from ssDNA. However, a relatively slow dissociation rate is required for the proposed deaminase-independent roadblock mechanism in which APOBEC3G binds the viral template strand and blocks reverse transcriptase- catalysed DNA elongation. Here, we show that APOBEC3G initially binds ssDNA with rapid on-off rates and subsequently converts to a slowly dissociating mode. In contrast, an oligomerization-deficient APOBEC3G mutant did not exhibit a slow off rate. We propose that catalytically active monomers or dimers slowly oligomerize on the viral genome and inhibit reverse transcription.

AB - The human APOBEC3 proteins are a family of DNA-editing enzymes that play an important role in the innate immune response against retroviruses and retrotransposons. APOBEC3G is a member of this family that inhibits HIV-1 replication in the absence of the viral infectivity factor Vif. Inhibition of HIV replication occurs by both deamination of viral single-stranded DNA and a deamination-independent mechanism. Efficient deamination requires rapid binding to and dissociation from ssDNA. However, a relatively slow dissociation rate is required for the proposed deaminase-independent roadblock mechanism in which APOBEC3G binds the viral template strand and blocks reverse transcriptase- catalysed DNA elongation. Here, we show that APOBEC3G initially binds ssDNA with rapid on-off rates and subsequently converts to a slowly dissociating mode. In contrast, an oligomerization-deficient APOBEC3G mutant did not exhibit a slow off rate. We propose that catalytically active monomers or dimers slowly oligomerize on the viral genome and inhibit reverse transcription.

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Oligomerization transforms human APOBEC3G from an efficient enzyme to a slowly dissociating nucleic acid-binding protein

Abstract

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