An insight into the dependence of the deamination rate of human APOBEC3F on the length of single-stranded DNA, which is affected by the concentrations of APOBEC3F and single-stranded DNA
被引:2
作者:
Wan, Li
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Kyoto Univ, Inst Adv Energy, Uji, Kyoto, Japan
Kyoto Univ, Grad Sch Energy Sci, Uji, Kyoto, JapanKyoto Univ, Inst Adv Energy, Uji, Kyoto, Japan
Wan, Li
[1
,2
]
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Kamba, Keisuke
[1
]
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Nagata, Takashi
[1
,2
]
Katahira, Masato
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机构:
Kyoto Univ, Inst Adv Energy, Uji, Kyoto, Japan
Kyoto Univ, Grad Sch Energy Sci, Uji, Kyoto, JapanKyoto Univ, Inst Adv Energy, Uji, Kyoto, Japan
Katahira, Masato
[1
,2
]
机构:
[1] Kyoto Univ, Inst Adv Energy, Uji, Kyoto, Japan
[2] Kyoto Univ, Grad Sch Energy Sci, Uji, Kyoto, Japan
Background: APOBEC3F (A3F), a member of the human APOBEC3 (A3) family of cytidine deaminases, acts as an anti-HIV-1 factor by deaminating deoxycytidine in the complementary DNA of the viral genome. A full understanding of the deamination behavior of A3F awaits further investigation. Methods: The real-time NMR method and uracil-DNA glycosylase assay were used to track the activities of the C-terminal domain (CTD) of A3F at different concentrations of A3F-CTD and ssDNA. The steady-state fluorescence anisotropy measurement was used to examine the binding between A3F-CTD and ssDNA with different lengths. The use of the A3F-CTD N214H mutant, having higher activity than the wild-type, facilitated the tracking of the reactions. Results: A3F-CTD was found to efficiently deaminate the target deoxycytidine in long ssDNA in lower ssDNA concentration conditions ([A3F-CTD] >> [ssDNA]), while the target deoxycytidine in short ssDNA is deaminated efficiently in higher ssDNA concentration conditions ([A3F-CTD] << [ssDNA]). This property is quite different from that of the previously studied A3 family member, A3B; the concentrations of the proteins and ssDNA had no effect. Conclusions: The concentrations of A3F-CTD and ssDNA substrates affect the ssDNA-length-dependence of deamination rate of the A3F-CTD. This unique property of A3F is rationally interpreted on the basis of its binding characteristics with ssDNA. General significance: The discovery of the unique property of A3F regarding the deamination rate deepens the understanding of its counteraction against HIV-1. Our strategy is applicable to investigate the other aspects of the A3 activities, such as those involved in the cancer development.
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Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
Aydin, Halil
Taylor, Matthew W.
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Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
Taylor, Matthew W.
Lee, Jeffrey E.
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Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Bishop, KN
Holmes, RK
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机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Holmes, RK
Sheehy, AM
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机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Sheehy, AM
Davidson, NO
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机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Davidson, NO
Cho, SJ
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机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Cho, SJ
Malim, MH
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Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, EnglandKings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
机构:
NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Chaipan, Chawaree
Smith, Jessica L.
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NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Smith, Jessica L.
Hu, Wei-Shau
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NCI, Viral Recombinat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Hu, Wei-Shau
Pathak, Vinay K.
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机构:
NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
机构:
Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
Aydin, Halil
Taylor, Matthew W.
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h-index: 0
机构:
Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
Taylor, Matthew W.
Lee, Jeffrey E.
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h-index: 0
机构:
Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, CanadaUniv Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON M5S 1A8, Canada
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Bishop, KN
Holmes, RK
论文数: 0引用数: 0
h-index: 0
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Holmes, RK
Sheehy, AM
论文数: 0引用数: 0
h-index: 0
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Sheehy, AM
Davidson, NO
论文数: 0引用数: 0
h-index: 0
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Davidson, NO
Cho, SJ
论文数: 0引用数: 0
h-index: 0
机构:Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
Cho, SJ
Malim, MH
论文数: 0引用数: 0
h-index: 0
机构:
Kings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, EnglandKings Coll London, Guys Kings & St Thomas Sch Med, Dept Infect Dis, London SE1 9RT, England
机构:
NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Chaipan, Chawaree
Smith, Jessica L.
论文数: 0引用数: 0
h-index: 0
机构:
NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Smith, Jessica L.
Hu, Wei-Shau
论文数: 0引用数: 0
h-index: 0
机构:
NCI, Viral Recombinat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA
Hu, Wei-Shau
Pathak, Vinay K.
论文数: 0引用数: 0
h-index: 0
机构:
NCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USANCI, Viral Mutat Sect, HIV Drug Resistance Program, Ctr Canc Res, Frederick, MD 21701 USA