Identification and functional analysis of the small membrane-associated protein pUL11 of avian infectious laryngotracheitis virus.
pUL11 is a highly conserved, small, acylated, membrane-associated tegument protein of herpesviruses. It is involved in final envelopment of nascent virions in the cytoplasm, although the precise mechanism is still unknown. By screening of mouse monoclonal antibodies (mAb) raised against purified particles of infectious laryngotracheitis virus (ILTV) of chickens (Veits et al., 2003a), we identified two mAb recognizing the 15 kDa UL11 protein (pUL11) of this avian alphaherpesvirus. These mAb permitted detection and precise localization of pUL11 in mature ILT virions, as well as in the cytoplasm of infected chicken cells by Western blot analyses, indirect immunofluorescence tests, and immunoelectron microscopy. For investigation of gene function UL11-deleted ILTV recombinants were generated. Like its homologues in several other alphaherpesviruses, ILTV-pUL11 was shown to be nonessential for productive virus replication. However, compared to wild-type and UL11 rescued ILTV the deletion mutants exhibited significantly reduced virus yields and moderately impaired spread in cell culture. In the absence of pUL11, electron microscopy of infected cells revealed accumulations of tegument proteins with nucleocapsids, and marked distortions of Golgi membranes in the cytoplasm, which obviously inhibited the formation of mature, enveloped virus particles. Taken together, our results demonstrate that pUL11 is relevant for secondary envelopment of ILTV, and confirm functional conservation of this protein in herpesviruses. The now available unique pUL11-specific mAb will help to further analyze this function, which is presumably mediated by physical interactions with other viral gene products, in cultured cells and in the natural animal host of ILTV.
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald - Insel Riems, Germany. walter.fuchs@fli.bund.de
Walter
W
Fuchs
Harald
H
Granzow
Jutta
J
Veits
Thomas C
TC
Mettenleiter
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002478:Cells, Cultured; D002645:Chickens; D017353:Gene Deletion; D001352:Herpesvirus 1, Gallid / Q000502:physiology*; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D015388:Organelles / Q000648:ultrastructure / Q000821:virology; D014764:Viral Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014771:Virion / Q000737:chemistry; D019065:Virus Assembly
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D008565:Membrane Proteins; D014764:Viral Proteins