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J. Biol. Chem., Vol. 276, Issue 25, 22265-22272, June 22, 2001
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From the Department of Medical Microbiology and Hygiene, Johannes
Gutenberg-Universität Mainz, D-55101 Mainz, Germany
The large (L) envelope protein of the hepatitis B
virus (HBV) has the peculiar capacity to form two transmembrane
topologies via an as yet uncharacterized process of partial
post-translational translocation of its pre-S domain across
membranes. In view of a current model that predicts an HBV-specific
channel generated during virion envelope assembly to enable pre-S
translocation, we have examined parameters influencing L topogenesis by
using protease protection analysis of wild-type and mutant L proteins synthesized in transfected cells. We demonstrate that contrary to expectation, all determinants, thought to be responsible for channel
formation, are dispensable for pre-S reorientation. In particular, we
observed that this process does not require (i) the helper function of
the HBV S (small) and M (middle) envelope proteins, (ii) covalent dimer
formation of envelope chains, or (iii) either of the three amphipathic
transmembrane segments of L. Rather, the most hydrophobic transmembrane
segment 2 of L was identified as a vital topogenic determinant,
essential and sufficient for post-translational pre-S translocation.
Cell fractionation studies revealed that pre-S refolding and thus the
dual topology of L is established at the endoplasmic reticulum (ER)
membrane rather than at a post-ER compartment as originally supposed.
Together our data provide evidence to suggest that the topological
reorientation of L is facilitated by a host cell transmembrane
transport machinery such as the ER translocon.
Dual Topology of the Hepatitis B Virus Large Envelope Protein
DETERMINANTS INFLUENCING POST-TRANSLATIONAL PRE-S
TRANSLOCATION*
*
This work was supported by Deutsche Forschungsgemeinschaft
Grant SFB 490-A1 (to R. P.). C. L. was supported by Graduiertenkolleg 194.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Institute for Medical
Microbiology and Hygiene, University of Mainz, Augustusplatz, D-55101
Mainz, Germany. Tel.: 0049-6131-393-6750; Fax: 0049-6131-393-2359; E-mail: prange@mail.uni-mainz.de.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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