Characterization of subcellular localization of duck enteritis virus UL51 protein.
BACKGROUND
Knowledge of the subcellular localization of a protein can provide useful insights about its function. While the subcellular localization of many alphaherpesvirus UL51 proteins has been well characterized, little is known about where duck enteritis virus (DEV) UL51 protein (pUL51) is targeted to. Thus, in this study, we investigated the subcellular localization and distribution of DEV pUL51 by computer aided analysis, as well as indirect immunofluorescence (IIF) and transmission immunoelectron microscopy (TIEM) approaches in DEV-infected cells.


RESULTS
The DEV UL51 gene product was identified as an approximate 34 kDa protein in DEV-infected cells analyzed by western blotting. Computer aided analysis suggested that DEV pUL51 is not targeted to the mitochondrial, extra-cellular or nucleus, but be targeted to the cytoplasmic in host cells, more specifically, palmitoylation of the pUL51 through the N-terminal cysteine at position 9 makes membrane association and Golgi localization possible. Using IIF analysis, we found that DEV pUL51 was first detected in a juxtanuclear region of DEV-infected cells at 9 h postinfection (p.i.), and then was detected widely distributed in the cytoplasm and especially was stronger in the juxtanuclear region from 12 to 60 h p.i. TIEM analysis revealed that DEV pUL51 was mainly associated with cytoplasmic virions and also with some membranous structure near the pUL51-specific immuno-labeling intracellular virion in the cytoplasmic vesicles; moreover, the pUL51 efficiently accumulated in the Golgi apparatus at first, and then was sent to the plasma membrane from the Golgi by some unknown mechanism.


CONCLUSIONS
In this work, we described the basic characteristics of pUL51 subcellular localization and distribution for the first time. From these results, we concluded that palmitoylation at the N-terminal cysteine, which is conserved in all alphaherpesvirus UL51 homologs, is required for its membrane association and Golgi localization, and the pUL51 mainly localized to the juxtanuclear region of DEV-infected cells, as well seemed to be incorporated into mature virions as a component of the tegument. The research will provide useful clues for DEV pUL51 functional analysis, and will be usefull for further understanding the localization properties of alphaherpesvirus UL51 homologs.
Shen
Chanjuan
C
Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Ya'an, Sichuan, PR China. vorber@163.com
Guo
Yufei
Y
Cheng
Anchun
A
Wang
Mingshu
M
Zhou
Yi
Y
Lin
Dan
D
Xin
Hongyi
H
Zhang
Na
N
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry; D022162:Cytoplasmic Vesicles / Q000737:chemistry; D004372:Ducks; D005347:Fibroblasts / Q000821:virology; D006056:Golgi Apparatus / Q000737:chemistry; D006564:Herpesviridae / Q000502:physiology*; D054878:Lipoylation; D008856:Microscopy, Fluorescence / Q000379:methods; D016253:Microscopy, Immunoelectron / Q000379:methods; D008970:Molecular Weight; D015388:Organelles / Q000737:chemistry*; D014764:Viral Proteins / Q000032:analysis* / Q000737:chemistry; D014779:Virus Replication
D014764:Viral Proteins