Human cytomegalovirus capsid assembly protein precursor (pUL80.5) interacts with itself and with the major capsid protein (pUL86) through two different domains

Wood, Lisa J.; Baxter, Michael K.; Plafker, Scott M.; Gibson, Wade

doi:10.1128/jvi.71.1.179-190.1997

Cited by 69 publications

(38 citation statements)

References 74 publications

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“…At least 11 amino acids at the extreme carboxyl terminus of the P22 scaffolding protein are required for procapsid assembly and binding to the coat protein (Parker et al, 1998). A similar requirement for about 15±25 residues at the carboxyl terminus has been described for the scaffolding proteins from the related viruses herpes simplex type I (HSV-1; Matusick-Kumar et al, 1995; and cytomegalovirus (CMV;Wood et al, 1997). The scaffolding proteins from all three of these species have a pattern of hydrophobic residues at the carboxyl terminus that strongly suggests that they form amphipathic helices (Cohen and Parry, 1990), and helicity in this domain has been shown to be required for scaffolding/major capsid protein interactions in HSV-1 (Hong et al, 1996).…”

Section: Introductionmentioning

confidence: 71%

“…For most viruses with T number Ͻ7, all of the information required for coat protein subunit conformational switching is contained within the subunits themselves. However, for viruses with T number Ն7, such as dsDNA phage (Casjens and Hendrix, 1988), herpesviruses (Hong et al, 1996;Wood et al, 1997), and adenoviruses (D'Halluin et al, 1978;Hasson et al, 1989Hasson et al, , 1992) the assistance of a scaffolding protein, which both directs the form determination and accelerates the assembly process, is required. Therefore, understanding of the assembly processes for these viruses requires insight into the mechanisms by which scaffolding proteins interact with the coat or capsid proteins.…”

Section: Discussionmentioning

confidence: 99%

“…The Salmonella typhimurium bacteriophage P22 assembles by a pathway that is typical of the doublestranded DNA (dsDNA) 2 phage (Hendrix, 1985;Casjens and Hendrix, 1988) and of related viruses such as herpesviruses (Rixon, 1993;Donaghy and Jupp, 1995;Hong et al, 1996;Newcomb et al, 1996;Trus et al, 1996;Wood et al, 1997) and adenoviruses (D'Halluin et al, 1978). The initial step in P22 assembly is the copolymerization of 420 molecules of the coat protein (gp5) into a Tϭ7 icosahedral shell termed a`procapsid'' (Casjens, 1979;Prasad et al, 1993;Thuman-Commike et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Electrostatic Interactions Drive Scaffolding/Coat Protein Binding and Procapsid Maturation in Bacteriophage P22

Parker

Prevelige

1998

Virology

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The first step in assembly of the bacteriophage P22 is the formation of a T=7 icosahedral "procapsid," the major components of which are the coat protein and an inner core composed of the scaffolding protein. Although not present in the mature virion, the scaffolding protein is required for procapsid assembly. Eleven amino-acid residues at the extreme carboxyl terminus of the scaffolding protein are required for binding to the coat protein, and upon deletion of these residues, approximately 20 additional residues become disordered. Sequence analysis and NMR data suggest that the 30 residues at the carboxyl terminus form a helix-loop-helix motif which is stabilized by interhelical hydrophobic interactions. This "coat protein recognition domain" presents an unusually high number of positively charged residues on one face, suggesting that electrostatic interactions between this domain and the coat protein may contribute to recognition and binding. We report here that high ionic strength (1 M NaCl) completely inhibited procapsid assembly in vitro. When scaffolding protein was added to empty procapsid "shells" of coat protein, 1 M NaCl partially inhibited the binding of scaffolding protein to the shells. This suggests that the positively charged coat protein recognition domain at the carboxyl terminus of the scaffolding protein binds to a negatively charged region on the coat protein. During DNA packaging, the scaffolding protein exits the procapsid; scaffolding protein exit is followed by the expansion of the procapsid into a mature capsid. Procapsid shells can be induced to undergo a similar expansion reaction in vitro by heating (45-70 degreesC); this process was also inhibited by 1 M NaCl. These results are consistent with a model in which negatively charged scaffold protein-binding domains in the coat proteins move apart during procapsid expansion; this relief of electrostatic repulsion could provide a driving force for expansion and subsequent maturation. High-salt concentrations would screen this repulsion, while packaging of DNA (a polyanion) in vivo may increase the instability of the procapsid enough to trigger its expansion.

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Section: Introductionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Electrostatic Interactions Drive Scaffolding/Coat Protein Binding and Procapsid Maturation in Bacteriophage P22

Parker

Prevelige

1998

Virology

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“…The outer domain is presumably the C-terminal moiety, since the C terminus of the precursor should interact with the surface shell (7,21,36,44,45,69,71). We tentatively suggest that the linker may be an ␣-helical coiled coil, about four heptads (4 nm) long, based on scanning the SCMV assembly protein sequence (68) with a coiled-coil detection algorithm (40) (unpublished results).…”

Section: Discussionmentioning

confidence: 99%

Capsid Structure of Simian Cytomegalovirus from Cryoelectron Microscopy: Evidence for Tegument Attachment Sites

Trus

Gibson

Cheng

et al. 1999

J Virol

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We have used cryoelectron microscopy and image reconstruction to study B-capsids recovered from both the nuclear and the cytoplasmic fractions of cells infected with simian cytomegalovirus (SCMV). SCMV, a representative betaherpesvirus, could thus be compared with the previously described B-capsids of the alphaherpesviruses, herpes simplex virus type 1 (HSV-1) and equine herpesvirus 1 (EHV-1), and of channel catfish virus, an evolutionarily remote herpesvirus. Nuclear B-capsid architecture is generally conserved with SCMV, but it is 4% larger in inner radius than HSV-1, implying that its ∼30% larger genome should be packed more tightly. Isolated SCMV B-capsids retain a relatively well preserved inner shell (or “small core”) of scaffolding-assembly protein, whose radial-density profile indicates that this protein is ∼16-nm long and consists of two domains connected by a low-density linker. As with HSV-1, the hexons but not the pentons of the major capsid protein (151 kDa) bind the smallest capsid protein (∼8 kDa). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed cytoplasmic B-capsid preparations to contain proteins similar in molecular weight to the basic phosphoprotein (∼119 kDa) and the matrix proteins (65 to 70 kDa). Micrographs revealed that these particles had variable amounts of surface-adherent material not present on nuclear B-capsids that we take to be tegument proteins. Cytoplasmic B-capsids were classified accordingly as lightly, moderately, or heavily tegumented. By comparing the three corresponding density maps with each other and with the nuclear B-capsid, two interactions were identified between putative tegument proteins and the capsid surface. One is between the major capsid protein and a protein estimated by electron microscopy to be 50 to 60 kDa; the other involves an elongated molecule estimated to be 100 to 120 kDa that is anchored on the triplexes, most likely on its dimer subunits. Candidates for the proteins bound at these sites are discussed. This first visualization of such linkages makes a step towards understanding the organization and functional rationale of the herpesvirus tegument.

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“…Immature capsids lack DNA but contain an abundant internal polypeptide, the assembly protein (AP), that is not found in the mature, DNA-containing particles (25,27,34,48,55). This protein is known to interact, through its carboxy-terminal domain, with the major capsid protein (32,69). This interaction is required for nuclear transport of the major capsid protein (47) and has also been proposed to act as a scaffold to facilitate the assembly of the capsid shell.…”

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confidence: 99%

The protease and the assembly protein of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8)

Ünal

Pray

Lagunoff

et al. 1997

J Virol

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A genomic clone encoding the protease (Pr) and the assembly protein (AP) of Kaposi's sarcoma-associated herpesvirus (KSHV) (also called human herpesvirus 8) has been isolated and sequenced. As with other herpesviruses, the Pr and AP coding regions are present within a single long open reading frame. The mature KSHV Pr and AP polypeptides are predicted to contain 230 and 283 residues, respectively. The amino acid sequence of KSHV Pr has 56% identity with that of herpesvirus saimiri, the most similar virus by phylogenetic comparison. Pr is expressed in infected human cells as a late viral gene product, as suggested by RNA analysis of KSHV-infected BCBL-1 cells. Expression of the Pr domain in Escherichia coli yields an enzymatically active species, as determined by cleavage of synthetic peptide substrates, while an active-site mutant of this same domain yields minimal proteolytic activity. Sequence comparisons with human cytomegalovirus (HCMV) Pr permitted the identification of the catalytic residues, Ser114, His46, and His134, based on the known structure of the HCMV enzyme. The amino acid sequences of the release site of KSHV Pr (Tyr-Leu-Lys-Ala*Ser-Leu-Ile-Pro) and the maturation site (Arg-Leu-Glu-Ala*Ser-Ser-Arg-Ser) show that the extended substrate binding pocket differs from that of other members of the family. The conservation of amino acids known to be involved in the dimer interface region of HCMV Pr suggests that KSHV Pr assembles in a similar fashion. These features of the viral protease provide opportunities to develop specific inhibitors of its enzymatic activity.

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Human cytomegalovirus capsid assembly protein precursor (pUL80.5) interacts with itself and with the major capsid protein (pUL86) through two different domains

Cited by 69 publications

References 74 publications

Electrostatic Interactions Drive Scaffolding/Coat Protein Binding and Procapsid Maturation in Bacteriophage P22

Electrostatic Interactions Drive Scaffolding/Coat Protein Binding and Procapsid Maturation in Bacteriophage P22

Capsid Structure of Simian Cytomegalovirus from Cryoelectron Microscopy: Evidence for Tegument Attachment Sites

The protease and the assembly protein of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8)

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