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Journal of Virology, June 1999, p. 4600-4610, Vol. 73, No. 6
Department of Microbiology and Immunology,
Wake Forest University School of Medicine, Winston-Salem, North
Carolina 27157-1064
Received 9 November 1998/Accepted 23 February 1999
A region in the carboxy terminus of the protein encoded by open
reading frame 6 in early region 4 (E4orf6) of adenovirus type 5 was
determined to be required for directing nuclear localization of the E1B
55-kDa protein and for efficient virus replication. A peptide
encompassing this region, corresponding to amino acids 239 through 255 of the E4orf6 protein, was analyzed by circular dichroism spectroscopy. The peptide showed evidence of
self-interaction and displayed the characteristic spectra of an
amphipathic
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
An Arginine-Faced Amphipathic Alpha Helix Is
Required for Adenovirus Type 5 E4orf6 Protein Function
helix in the helix-stabilizing solvent
trifluoroethanol. Disrupting the integrity of this
helix in the
E4orf6 protein by proline substitutions or by removing amino acids 241 through 250 abolished its ability to direct the E1B 55-kDa protein to
the nucleus when both proteins were transiently expressed in HeLa
cells. Expression of E4orf6 variants that failed to direct nuclear
localization of the E1B 55-kDa protein failed to enhance replication of
the E4 mutant virus, dl1014, whereas expression of the
wild-type E4orf6 protein restored growth of dl1014 to
near-wild-type levels. These results suggest that the E4orf6 protein
contains an arginine-faced, amphipathic
helix that is critical for
a functional interaction with the E1B 55-kDa protein in the cell and
for the function of the E4orf6 protein during a lytic infection.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Wake Forest University School of Medicine, Bowman Gray Campus, Winston-Salem, NC 27157-1064. Phone: (336) 716-9332. Fax: (336) 716-9928. E-mail: ornelles{at}wfubmc.edu.
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