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Journal of Virology, July 2004, p. 7717-7726, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7717-7726.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Human Cytomegalovirus Carries a Cell-Derived Phospholipase A2 Required for Infectivity

Cuider Allal,1 Claire Buisson-Brenac,1 Vincent Marion,1 Clotilde Claudel-Renard,2 Thomas Faraut,2 Paola Dal Monte,3 Daniel Streblow,4 Michel Record,1,{dagger}* and Jean-Luc Davignon1,{dagger}*

INSERM U563, Toulouse, CPTP, Institut Claude de Préval, IFR30, CHU Purpan, 31059 Toulouse Cedex,1 Laboratoire de Génétique Cellulaire, INRA, BP27, 31326 Castanet Tolosan, France,2 Department of Clinical and Experimental Medicine, Division of Microbiology, University of Bologna, 40138 Bologna, Italy,3 Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland, Oregon 972014

Received 11 February 2004/ Accepted 15 March 2004

Human cytomegalovirus (HCMV) is known to carry host cell-derived proteins and mRNAs whose role in cell infection is not understood. We have identified a phospholipase A2 (PLA2) activity borne by HCMV by using an assay based on the hydrolysis of fluorescent phosphatidylcholine. This activity was found in all virus strains analyzed and in purified strains. It was calcium dependent and was sensitive to inhibitors of cytosolic PLA2 (cPLA2) but not to inhibitors of soluble PLA2 or calcium-independent PLA2. No other phospholipase activity was detected in the virus. Purified virus was found to contain human cellular cPLA2{alpha}, as detected by monoclonal antibody. No homology with PLA2 was found in the genome of HCMV, indicating that HCMV does not code for a PLA2. Decreased de novo expression of immediate-early proteins 1 and 2 (IE1 and IE2), tegument phosphoprotein pp65, and virus production was observed when HCMV was treated with inhibitors of cPLA2. Cell entry of HCMV was not altered by those inhibitors, suggesting the action of cPLA2 was postentry. Together, our results indicate a selective sorting of a cell-derived cPLA2 during HCMV maturation, which is further required for infectivity.

* Corresponding author. Mailing address for M. Record: INSERM U563, Toulouse, CPTP, Institut Claude de Préval, IFR30, CHU Purpan, 31059 Toulouse Cedex, France. Phone: 33 561 77 94 11. Fax: 33 561 77 94 01. E-mail: record{at}cict.fr. Mailing address for J.-L. Davignon: INSERM U563, Toulouse, CPTP, Institut Claude de Préval, IFR30, CHU Purpan 31059 Toulouse Cedex, France. Phone: 33 562 74 83 69. Fax: 33 562 74 83 86. E-mail: davignon{at}toulouse.inserm.fr.

{dagger} M.R. and J.-L.D. contributed equally to the supervision of this work.

Journal of Virology, July 2004, p. 7717-7726, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7717-7726.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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