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Journal of Virology, February 1999, p. 1293-1301, Vol. 73, No. 2
Tsukuba Life Science Center,
Received 20 July 1998/Accepted 31 October 1998
The cytoplasmic domain of an envelope transmembrane glycoprotein
(gp30) of bovine leukemia virus (BLV) has two overlapping copies of the
(YXXL)2 motif. The N-terminal motif has been implicated in
in vitro signal transduction pathways from the external to the
intracellular compartment and is also involved in infection and
maintenance of high viral loads in sheep that have been experimentally infected with BLV. To determine the role of YXXL sequences in the
replication of BLV in vitro, we changed the tyrosine or leucine residues of the N-terminal motif in an infectious molecular clone of
BLV, pBLV-IF, to alanine to produce mutated proviruses designated Y487A, L490A, Y498A, L501A, and Y487/498A. Transient transfection of
African green monkey kidney COS-1 cells with proviral DNAs that encoded
wild-type and mutant sequences revealed that all of the mutated
proviral DNAs synthesized mature envelope proteins and released virus
particles into the growth medium. However, serial passages of fetal
lamb kidney (FLK) cells, which are sensitive to infection with BLV,
after transient transfection revealed that mutation of a second
tyrosine residue in the N-terminal motif completely prevented the
propagation of the virus. Similarly, Y498A and Y487/498A mutant BLV
that was produced by the stably transfected COS-1 cells exhibited
significantly reduced levels of cell-free virion-mediated transmission.
Analysis of the protein compositions of mutant viruses demonstrated
that lower levels of envelope protein were incorporated by two of the
mutant virions than by wild-type and other mutant virions. Furthermore,
a mutation of a second tyrosine residue decreased the specific binding
of BLV particles to FLK cells and the capacity for viral penetration. Our data indicate that the YXXL sequences play critical roles in both
viral entry and the incorporation of viral envelope protein into the
virion during the life cycle of BLV.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The YXXL Sequences of a Transmembrane Protein of Bovine Leukemia
Virus Are Required for Viral Entry and Incorporation of Viral
Envelope Protein into Virions
*
Corresponding author. Mailing address: Tsukuba Life
Science Center, The Institute of Physical and Chemical Research
(RIKEN), 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan. Phone: 81 298 36 3522. Fax: 81 298 36 9050. E-mail:
aida{at}rtc.riken.go.jp.
Journal of Virology, February 1999, p. 1293-1301, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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