HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity

Qingqing Chai; Sunan Li; Morgan K. Collins; Rongrong Li; Iqbal Ahmad; Silas F. Johnson; Dylan A. Frabutt; Zhichang Yang; Xiaojing Shen; Liangliang Sun; Jian Hu; Judd F. Hultquist; B. Matija Peterlin; Yong Hui Zheng

doi:10.1016/j.celrep.2021.109514

HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity

Qingqing Chai, Sunan Li, Morgan K. Collins, Rongrong Li, Iqbal Ahmad, Silas F. Johnson, Dylan A. Frabutt, Zhichang Yang, Xiaojing Shen, Liangliang Sun, Jian Hu, Judd F. Hultquist, B. Matija Peterlin, Yong Hui Zheng^*

^*Corresponding author for this work

Medicine, Infectious Diseases Division

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

10 Scopus citations

Abstract

HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.

Original language	English (US)
Article number	109514
Journal	Cell reports
Volume	36
Issue number	6
DOIs	https://doi.org/10.1016/j.celrep.2021.109514
State	Published - Aug 10 2021

Funding

We thank Heinrich Gottlinger, Qintong Li, Jennifer Doudna, and the NIH AIDS Research and Reference Reagent Program for providing reagents. The graphical abstract and the model in Figure 7D were created with BioRender. Y.-H.Z. is supported by a grant from the National Institutes of Health (AI145504). Q.C. S.L. M.K.C. R.L. I.A. and D.A.F. performed all experiments except the MS analyses. Z.Y. X.S. and L.S. conducted all MS analyses. J.H. supported SERINC5 protein purification. J.F.H. supported silencing CCNK in HEK293T and Jurkat cells. S.F.J. created the graphical abstract and the model in Figure 7D. B.M.P. edited the manuscript. Y.-H.Z. designed this study and wrote the manuscript with input from all co-authors. The authors declare no competing interests. We thank Heinrich Gottlinger, Qintong Li, Jennifer Doudna, and the NIH AIDS Research and Reference Reagent Program for providing reagents. The graphical abstract and the model in Figure 7 D were created with BioRender. Y.-H.Z. is supported by a grant from the National Institutes of Health ( AI145504 ).

Keywords

AP-2
CDK
HIV-1
Nef
SERINC5
cyclin
endocytosis
intrinsic immunity
restriction factors
serine phosphorylation

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.celrep.2021.109514

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title = "HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity",

abstract = "HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.",

keywords = "AP-2, CDK, HIV-1, Nef, SERINC5, cyclin, endocytosis, intrinsic immunity, restriction factors, serine phosphorylation",

author = "Qingqing Chai and Sunan Li and Collins, {Morgan K.} and Rongrong Li and Iqbal Ahmad and Johnson, {Silas F.} and Frabutt, {Dylan A.} and Zhichang Yang and Xiaojing Shen and Liangliang Sun and Jian Hu and Hultquist, {Judd F.} and Peterlin, {B. Matija} and Zheng, {Yong Hui}",

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doi = "10.1016/j.celrep.2021.109514",

language = "English (US)",

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journal = "Cell reports",

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T1 - HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity

AU - Chai, Qingqing

AU - Li, Sunan

AU - Collins, Morgan K.

AU - Li, Rongrong

AU - Ahmad, Iqbal

AU - Johnson, Silas F.

AU - Frabutt, Dylan A.

AU - Yang, Zhichang

AU - Shen, Xiaojing

AU - Sun, Liangliang

AU - Hu, Jian

AU - Hultquist, Judd F.

AU - Peterlin, B. Matija

AU - Zheng, Yong Hui

PY - 2021/8/10

Y1 - 2021/8/10

N2 - HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.

AB - HIV-1-negative factor (Nef) protein antagonizes serine incorporator 5 (SERINC5) by redirecting this potent restriction factor to the endosomes and lysosomes for degradation. However, the precise mechanism remains unclear. Using affinity purification/mass spectrometry, we identify cyclin K (CycK) and cyclin-dependent kinase 13 (CDK13) as a Nef-associated kinase complex. CycK/CDK13 phosphorylates the serine at position 360 (S360) in SERINC5, which is required for Nef downregulation of SERINC5 from the cell surface and its counteractivity of the SERINC5 antiviral activity. To understand the role of S360 phosphorylation, we generate chimeric proteins between CD8 and SERINC5 to study their response to Nef. Nef not only downregulates but, importantly, also binds to this chimera in an S360-dependent manner. Thus, S360 phosphorylation increases interactions between Nef and SERINC5 and initiates the destruction of SERINC5 by the endocytic machinery.

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KW - restriction factors

KW - serine phosphorylation

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HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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