Isoprenylation masks a conformational epitope and enhances trans-dominant inhibitory function of the large hepatitis delta antigen.
Hepatitis delta antigen (HDAg) consists of two species, large (LHDAg) and small (SHDAg), which are identical in sequence except that the large form contains 19 extra amino acids at the C terminus. The large form is prenylated on the Cxxx motif. The small form can trans activate HDV RNA replication, while the large form inhibits it. To determine the molecular basis for their differential functions, we examined the effects of prenylation on the conformation and function of HDAg. We show that the presence of prenylates masks a conformational epitope which is present in SHDAg but hidden in wild-type LHDAg; this epitope becomes exposed in all of the nonprenylated mutant LHDAgs. Prenylation also plays a major role in conferring the trans-dominant negative inhibitory activity of LHDAg, since the loss of prenylation in LHDAg reduced its inhibitory activity. The primary amino acids of the C-terminal sequence also contributed to the maintenance of the HDAg protein conformation; a prenylated LHDAg mutant with a five-amino-acid deletion had an exposed C-terminal epitope. By examining LHDAg mutants which have deletions of various extents of C-terminal sequence, with or without the prenylation motif, we have further shown that all of the prenylated mutants have much higher levels of trans-dominant suppressor activities than do the corresponding nonprenylated mutants. Surprisingly, a few nonprenylated LHDAg mutants were able to trans activate HDV RNA replication, while all of the prenylated ones lost this function. These results suggest that isoprenylates cause the masking of a conformational epitope of HDAg and that conformational differences between the large and small HDAgs account for the differences in their trans-activating and trans-dominant inhibitory biological activities.
Hwang
S B
SB
Howard Hughes Medical Institute, University of Southern California School of Medicine, Los Angeles 90033-1054.
Lai
M M
MM
D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D004252:DNA Mutational Analysis; D003673:Defective Viruses / Q000235:genetics* / Q000276:immunology; D000939:Epitopes; D015967:Gene Expression Regulation, Viral; D003698:Hepatitis Delta Virus / Q000235:genetics* / Q000276:immunology; D035923:Hepatitis delta Antigens; D011487:Protein Conformation; D017368:Protein Prenylation; D013329:Structure-Activity Relationship; D013489:Suppression, Genetic; D015533:Transcriptional Activation
D000956:Antigens, Viral; D000939:Epitopes; D035923:Hepatitis delta Antigens; C442244:hepatitis delta virus large antigen