title: A novel DCAF1-binding motif required for Vpx-mediated degradation of nuclear SAMHD1 and Vpr-induced G2 arrest.
abstract: HIV-2 and closely related SIV Vpx proteins are essential for viral replication in macrophages and dendritic cells. Vpx hijacks DCAF1-DDB1-Cul4 E3 ubiquitin ligase to promote viral replication. DCAF1 is essential for cell proliferation and embryonic development and is responsible for the polyubiquitination of poorly defined cellular proteins. How substrate receptors recruit the DCAF1-containing E3 ubiquitin ligase to induce protein degradation is still poorly understood. Here we identify a highly conserved motif (Wx4Φx2Φx3AΦxH) that is present in diverse Vpx and Vpr proteins of primate lentiviruses. We demonstrate that the Wx4Φx2Φx3AΦxH motif in SIVmac Vpx is required for both the Vpx-DCAF1 interaction and/or Vpx-mediated degradation of SAMHD1. DCAF1-binding defective Vpx mutants also have impaired ability to promote SIVΔVpx virus infection of myeloid cells. Critical amino acids in the Wx4Φx2Φx3AΦxH motif of SIV Vpx that are important for DCAF1 interaction maintained the ability to bind SAMHD1, indicating that the DCAF1 and SAMHD1 interactions involve distinctive interfaces in Vpx. Surprisingly, VpxW24A mutant proteins that were still capable of binding DCAF1 and SAMHD1 lost the ability to induce SAMHD1 degradation, suggesting that Vpx is not a simple linker between the DCAF1-DDB1-Cul4 E3 ubiquitin ligase and its substrate, SAMHD1.VpxW24A maintained the ability to accumulate in the nucleus despite the fact that nuclear, but not cytoplasmic, mutant forms of SAMHD1 were more sensitive to Vpx-mediated degradation. The Wx4Φx2Φx3AΦxH motif in HIV-1 Vpr is also required for the Vpr-DCAF1 interaction and Vpr-induced G2 cell cycle arrest. Thus, our data reveal previously unrecognized functional interactions involved in the assembly of virally hijacked DCAF1-DDB1-based E3 ubiquitin ligase complex.
authors: First Hospital of Jilin University, Institute of Virology and AIDS Research, Changchun, Jilin Province, China.
authors: Wei
authors: W
authors: Wei
authors: Haoran
authors: H
authors: Guo
authors: Xue
authors: X
authors: Han
authors: Xianjun
authors: X
authors: Liu
authors: Xiaohong
authors: X
authors: Zhou
authors: Wenyan
authors: W
authors: Zhang
authors: Xiao-Fang
authors: XF
authors: Yu
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D002453:Cell Cycle; D002460:Cell Line; D017124:Conserved Sequence; D016336:Gene Products, vpr / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism*; D015302:Simian Immunodeficiency Virus / Q000472:pathogenicity* / Q000502:physiology; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication; D054325:vpr Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D016336:Gene Products, vpr; C513059:VPX protein, Simian immunodeficiency virus; D054334:Viral Regulatory and Accessory Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; C514912:vpr protein, Human immunodeficiency virus 1; D020559:Monomeric GTP-Binding Proteins