Hydrophobic cluster analysis predicts an amino-terminal domain of varicella-zoster virus open reading frame 10 required for transcriptional activation.
Varicella-zoster virus open reading frame 10 (ORF10) protein, the homolog of the herpes simplex virus protein VP16, can transactivate immediate-early promoters from both viruses. A protein sequence comparison procedure termed hydrophobic cluster analysis was used to identify a motif centered at Phe-28, near the amino terminus of ORF10, that strongly resembles the sequence of the activating domain surrounding Phe-442 of VP16. With a series of GAL4-ORF10 fusion proteins, we mapped the ORF10 transcriptional-activation domain to the amino-terminal region (aa 5-79). Extensive mutagenesis of Phe-28 in GAL4-ORF10 fusion proteins demonstrated the importance of an aromatic or bulky hydrophobic amino acid at this position, as shown previously for Phe-442 of VP16. Transactivation by the native ORF10 protein was abolished when Phe-28 was replaced by Ala. Similar amino-terminal domains were identified in the VP16 homologs of other alphaherpesviruses. Hydrophobic cluster analysis correctly predicted activation domains of ORF10 and VP16 that share critical characteristics of a distinctive subclass of acidic activation domains.
Moriuchi
H
H
Medical Virology Section, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892, USA.
Moriuchi
M
M
Pichyangkura
R
R
Triezenberg
S J
SJ
Straus
S E
SE
Cohen
J I
JI
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D002522:Chlorocebus aethiops; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000096:biosynthesis; D017405:Herpes Simplex Virus Protein Vmw65 / Q000737:chemistry; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D010649:Phenylalanine; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D014157:Transcription Factors; D015533:Transcriptional Activation; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D017405:Herpes Simplex Virus Protein Vmw65; C090372:ORF 10 protein, varicella-zoster virus; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014764:Viral Proteins; D010649:Phenylalanine; D015500:Chloramphenicol O-Acetyltransferase