"We report that the transcript start site of UL3 mRNA isolated from HSV-1 infected cells maps to a position downstream of the predicted translation start site."
Constructed recombinant virus CB8116
Site mutation at predicted UL-3 transcript start site
"UL3 protein translation initiates at the second in-frame start codon of the UL3 ORF."
"UL-3 encodes a 224 amino acid protein"
Markovitz NS (Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD) (11 Jul 2007). "The HSV-1 UL3 transcript starts within the UL3 ORF and encodes a 224 amino acid protein". Journal of Virology. PMID17626086.{{cite journal}}: CS1 maint: multiple names: authors list (link)
pUL-3 Localizes to the nucleolus 4 to 6 hours post infection (h.p.i.)
4 h.p.i. Localization to nucleosomes has begun
5 h.p.i. Accumulations at nucleosomes has grown
6 h.p.i. Intranuclear "donut" formation of pUL-3
Colocalizes with ICP8 coincidentally with donut formation/6 h.p.i.
Nucleolar localization requies amino acids 100 through 164*
Putative Nuclear Locating Sequence (1 of 2) predicted at aa 143-147.
M100/164-GFP (Green Flouresecent Protein) mutant UL3 localized to the nucleolus.
Putative phosphorylation site (1 of 2) in amino acids in 1-99*
Yamada H, Jiang YM, Zhu HY, Inagaki-Ohara K, Nishiyama Y (
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Japan) (Aug 1999). "Nucleolar localization of the UL3 protein of herpes simplex virus type 2". The Journal of General Virology. 80 (8): 2157-64. PMID10466815. {{cite journal}}: line feed character in |author= at position 59 (help)CS1 maint: multiple names: authors list (link)
Sequence anaysis predictions on UL-3 ORF
to contain an N-glycosylation site
to be a glycoprotein
UL-3 expressed in baculovirus system
Four electrophoretic bands:
Two major bands at 30kDa and 31kDa
Two minor bands at 29kDa and 33kDa
33kDa form contained marked 32P isotope, after 32P labelling
"None of the expressed UL3 protein species were susceptible to tunicamycin treatment, suggesting that they were not N-linked glycosylated."
Cell fractionation showed localization in the cytoplasm and nucleus but not cell membrane, again suggesting a lack of N-linked glycosylation.
Mouse antibodies v. pUL-3 created by vaccination with baculovirus UL-3 proteins
Reacted with two (2) HSV-1 proteins
27kDa and 33kDa forms
"presumably represent the unphosphorylated and phosphorylated forms of UL3"
PMID8879142 (1996, Cedars-Sanai Medical Center Research Institute)
Predicted molecular weight 29681 Da.
HSV-2 UL-3 75% homologous to HSV-1.
Three electrophoretic bands: 28000, 30500, 33000 Da.
30500 and 33000 Da forms contained marked 32P isotope after 32P labelling
30500 (but not 33000) Da form convertable to 28000 form via alkaline phosphatase
Immunohistochemical cell staining to locate protein
Early stage perinuclear association
Late stage associated with nucleus in discrete particles
PMID8337818 (1993, Department of Molecular Biology, University of Medicine and Dentistry of New Jersey, Stratford )
* Amino acids numbered from first start codon, pre-2007 numbering.
