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Journal of Virology, January 2005, p. 225-233, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.225-233.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Carboxyl-Terminal Region of Human Cytomegalovirus IE1491aa Contains an Acidic Domain That Plays a Regulatory Role and a Chromatin-Tethering Domain That Is Dispensable during Viral Replication

Jens Reinhardt,1,{dagger} Geoffrey B. Smith,1 Christopher T. Himmelheber,2 Jane Azizkhan-Clifford,2 and Edward S. Mocarski1*

Department of Microbiology & Immunology, Stanford University School of Medicine, Stanford, California,1 Department of Biochemistry, Drexel University College of Medicine, Philadelphia, Pennsylvania2

Received 4 June 2004/ Accepted 30 August 2004

The human cytomegalovirus major immediate-early ({alpha}) protein IE1491aa plays an important role in controlling viral gene expression at low multiplicities of infection. With a transient complementation assay, full-length IE1491aa enhanced the growth of ie1 mutant virus CR208 20-fold better than a deletion mutant lacking 71 carboxyl-terminal amino acids (IE11-420aa). A 16-amino-acid domain between amino acids 476 and 491 was both necessary and sufficient for chromatin-tethering activity; however, this domain was completely dispensable for complementation of CR208 replication. The proximal 55-amino-acid acidic domain (amino acids 421 to 475) was found to be most important for function. A deletion mutant lacking only this domain retained chromatin-tethering activity but failed to complement mutant virus. Interestingly, serine phosphorylation (at amino acids 399, 402, 406, 423, 428, 431, 448, 451, and 455) was not required for complementation. These results show that IE1491aa is composed of at least two domains that support replication, a region located between amino acids 1 and 399 that complements ie1 mutant virus replication to low levels and an acidic domain between amino acids 421 and 479 that dramatically enhances complementation.

* Corresponding author. Mailing address: Department of Microbiology & Immunology, Fairchild Science Building, 299 Campus Dr., Stanford University School of Medicine, Stanford, CA 94305-5124. Phone: (650) 723-6435. Fax: (650) 723-1606. E-mail: mocarski{at}stanford.edu.

{dagger} Present address: Station Biologique, Roscoff Cedex, France.

Journal of Virology, January 2005, p. 225-233, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.225-233.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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