Article Figures & Data
Figures
- Fig. 1.
Nomenclature and amino acid sequences of mutant DHBV core proteins. WT, wild-type core protein; 1N to 25N, mutated core proteins. Insertions are depicted in boldface letters.
- Fig. 2.
Southern blot analysis of viral DNA generated in mutant nucleocapsids. Nucleocapsid-associated viral DNA was isolated from cotransfected LMH cells after treatment with micrococcal nuclease. DH, viral DNA isolated from infected primary duck hepatocytes; 1N to 6N, modified core proteins (for nomenclature, see Fig. 1); WT, wild-type core protein; RC, relaxed-circular DNA; DL, double-stranded linear DNA; SS, single-stranded DNA; M, 3.0-kbp linear DHBV monomer.
- Fig. 3.
Apparent effect of size and charge of N-terminal insertions on viral DNA maturation. Results from Southern blot analysis are presented. Cytoplasmic extracts were treated with micrococcal nuclease prior to isolation of nucleocapsid-associated DNA. For nomenclature, see Fig. 1. The net charge of the respective insertion is indicated at the bottom. RC, relaxed-circular DNA; DL, double-stranded linear DNA; SS, single-stranded DNA; M, 3.0-kbp linear DHBV monomer.
- Fig. 4.
Endogenous polymerase activity in mutant nucleocapsids. Core particles were prepared from cotransfected LMH cells and incubated with radiolabeled nucleotides. Aliquots were processed directly (−MN) or after incubation with micrococcal nuclease (+MN). n.t., nontransfected LMH cells. For the nomenclature of the respective mutants, see Fig. 1. RC, relaxed-circular DNA.
- Fig. 5.
Effect of micrococcal nuclease digestion on the phenotype of N-terminally extended DHBV core proteins. Results from Southern blot analysis are presented. +MN, cytoplasmic extracts from cotransfected LMH cells treated with micrococcal nuclease prior to extraction of nucleocapsid-associated viral DNA. −MN, omission of micrococcal nuclease treatment. Instead of micrococcal nuclease, purified DNA samples were treated with the restriction enzymeDpnI, which selectively cuts methylated, transfected plasmid DNA but not the nonmethylated viral DNA. For the nomenclature of the respective mutants, see Fig. 1. WT, wild-type core protein; RC, relaxed-circular DNA; SS, single-stranded DNA; PL,DpnI fragment of transfected plasmid DNA.
- Fig. 6.
Effect of micrococcal nuclease digestion on the phenotype of C-terminally truncated DHBV core proteins. Results from Southern blot analysis are presented. For methods, see the legend to Fig. 5. +MN, treatment with micrococcal nuclease, but noDpnI digestion. −MN, no micrococcal nuclease treatment, but digestion with DpnI. WT, wild-type DHBV core protein; 239 and 243, C-terminally truncated core mutants according to Yu and Summers (31); RC, relaxed-circular DNA; DL, double-stranded linear DNA; SS, single-stranded DNA. Single-stranded DNA was visible on the right panel (−MN) upon longer exposure (not shown). PL, DpnI fragment of transfected plasmid DNA. M, 3.0-kbp linear DHBV monomer.
