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

Cell Rep. 2021 Aug 10;36(6):109514. doi: 10.1016/j.celrep.2021.109514.

Authors

Affiliations

¹ Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA.
² Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Harbin, China.
³ Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA; Department of Biology, Hillsdale College, Hillsdale, MI, USA.
⁴ Department of Chemistry, Michigan State University, East Lansing, MI, USA.
⁵ Department of Chemistry, Michigan State University, East Lansing, MI, USA; Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA.
⁶ Division of Infectious Diseases, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
⁷ Department of Medicine, University of California, San Francisco, San Francisco, CA, USA; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA, USA.
⁸ Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Harbin, China; Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA. Electronic address: zhengyo@msu.edu.

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.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Sequence
CDC2 Protein Kinase / metabolism*
Cyclins / metabolism*
Down-Regulation
HEK293 Cells
HIV Infections / metabolism
HIV Infections / virology*
HIV-1 / pathogenicity*
Humans
Jurkat Cells
Mass Spectrometry
Membrane Proteins / chemistry
Membrane Proteins / metabolism*
Peptides / chemistry
Peptides / metabolism
Phosphorylation
Phosphoserine / metabolism
Protein Binding
Proteomics
Recombinant Fusion Proteins / metabolism
nef Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

CCNK protein, human
Cyclins
Membrane Proteins
Peptides
Recombinant Fusion Proteins
SERINC5 protein, human
nef Gene Products, Human Immunodeficiency Virus
Phosphoserine
CDC2 Protein Kinase
CDK13 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding