Ring finger protein ZIN interacts with human immunodeficiency virus type 1 Vif.
Virion infectivity factor (Vif) protein of human immunodeficiency virus type 1 (HIV-1) is essential for the productive infection of primary human CD4 T lymphocytes and macrophages. Vif overcomes the HIV-inhibitory effects of cellular factor APOBEC3G, which has cytidine deaminase activity. We previously reported the isolation of a Vif-interacting ring finger protein, Triad 3, from a human leukocyte cDNA library, using the yeast two-hybrid system. The full-length cellular protein homologue of Triad 3 has been recently identified as the zinc finger protein inhibiting NF-kappaB (ZIN). Sequence analysis indicates that Triad 3 protein contains all four major ring-like motifs of ZIN. We report here that ZIN binds to purified Vif in vitro and that Triad 3/ZIN interacts with HIV-1 Vif in transfected human 293T cells, as demonstrated by coimmunoprecipitation. To test the biological relevance of this interaction, we produced infectious HIV-1 NL4.3 in the presence or absence of cotransfected ZIN. HIV-1 NL4.3 virus stocks produced in the presence of exogenously expressed ZIN were twofold less infectious in a single-cycle infectivity assay than virus produced in the absence of exogenous ZIN. It was further shown that cells infected with HIV NL4.3 virus stocks produced in the presence of exogenously expressed ZIN were impaired in viral DNA synthesis by twofold. The impairment in viral reverse transcription and the reduction in single-cycle viral infectivity were both shown to be dependent on the presence of Vif in the virus producer cells. The possible mechanisms by which ZIN interferes with the early events of HIV-1 replication are discussed.
Feng
Feng
F
Australian Centre for Hepatitis and HIV Virology Research, Infectious Disease Laboratories, Institute of Medical and Veterinary Science, Frome Rd., Adelaide, Australia 5000.
Davis
Adam
A
Lake
Julie-Anne
JA
Carr
Jill
J
Xia
Wei
W
Burrell
Christopher
C
Li
Peng
P
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D017353:Gene Deletion; D005786:Gene Expression Regulation; D016346:Gene Products, vif / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic; D044767:Ubiquitin-Protein Ligases; D014779:Virus Replication / Q000502:physiology; D016335:Zinc Fingers; D054320:vif Gene Products, Human Immunodeficiency Virus
D002352:Carrier Proteins; D004279:DNA, Viral; D016346:Gene Products, vif; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D054320:vif Gene Products, Human Immunodeficiency Virus; C458527:RNF216 protein, human; D044767:Ubiquitin-Protein Ligases