Genetic analysis of temperature-sensitive mutants of HSV-1: The combined use of complementation and physical mapping for cistron assignment

Weller, Sandra; Aschman, Doris P.; Sacks, Wendy R.; Coen, Donald M.; Schaffer, Priscilla A.

doi:10.1016/0042-6822(83)90084-3

Cited by 99 publications

(63 citation statements)

References 34 publications

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“…Our conclusion that the previously mapped tsQ26 mutation (23) lies in the gH gene shows that gH is essential, and the finding that the gH gene has counterparts in VZV and in EBV, which is not the case for several of the other HSV glycoproteins, could also argue a basic functional role for gH and its homologues.…”

Section: Methodsmentioning

confidence: 89%

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DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus

McGeoch

Davison

1986

Nucl Acids Res

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We have determined the sequence of herpes simplex virus type 1 DNA around the previously mapped location of sequences encoding an epitope of glycoprotein gH, and have deduced the structure of the gH gene and the amino acid sequence of gH. The unprocessed polypeptide is predicted to contain 838 amino acids, and to possess an N-terminal signal sequence and a C-terminal transmembrane sequence. Temperature-sensitive mutant tsQ26 maps within the predicted gH coding sequence. Homologous genes were identified in the genomes of two other herpesviruses, namely varicella-zoster virus and Epstein-Barr virus.

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Section: Methodsmentioning

confidence: 89%

“…In 1983, mapping of the temperature-sensitive mutation in HSV-1 strain KOS tsQ26 to a small region adjacent to the TK gene was reported (23). On the sequence listing of Figure 2, (30).…”

Section: Methodsmentioning

confidence: 99%

DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus

McGeoch

Davison

1986

Nucl Acids Res

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“…Identification of a mutation in tsQ26. The diagram shows the position of the gH and thymidine kinase (tk) coding sequences within the BglII m restriction fragment of HSV-1, and the location of the EcoRI and PvuII fragments that rescue tsQ26 (Weller et al, 1983). The gH coding sequence of strain KOS from the PvuII site to the EcoRI site is shown.…”

Section: ~Q26mentioning

confidence: 99%

“…The lesion in tsQ26 has been mapped by marker rescue in an EcoRI to PvuII fragment within map units 0.301 to 0.304 (Weller et al, 1983). To confirm the existence of a mutation in gH of tsQ26 the BglII m restriction fragments (map units 0-282 to 0.312) from DNA of tsQ26 and its parent KOS were cloned into the BglII site of pKC7 and the recombinant plasmids were digested with EcoRI and PvuII.…”

Section: Tsq26 Contains An Amino Acid Substitution In Ghmentioning

confidence: 99%

“…Chu et al (1979) isolated a temperature-sensitive (ts) mutant, tsQ26, from the parental strain KOS of HSV-1 and preliminary characterization of this mutant suggested a late defect since a normal pattern of protein synthesis was observed at the non-permissive temperature. Marker rescue experiments located the genetic lesion of tsQ26 in an EcoRI to PvulI fragment corresponding to map units 0.301 to 0.304 (Weller et al, 1983). As noted by McGeoch & Davison (1986) this fragment lies within the coding sequence of gH.…”

Section: Introductionmentioning

confidence: 99%

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Excretion of Non-infectious Virus Particles Lacking Glycoprotein H by a Temperature-sensitive Mutant of Herpes Simplex Virus Type 1: Evidence that gH Is Essential for Virion Infectivity

Desai

Schaffer

Minson

1988

Journal of General Virology

170

130

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SUMMARYA temperature-sensitive mutant of herpes simplex virus type 1, tsQ26, was shown to contain an amino acid substitution in glycoprotein H (gH). The mutant entered cells efficiently at the non-permissive temperature and replicated to give nearly normal yields of intracellular infectivity. The intracellular virions contained, predominantly, an immature form ofgH and no gH was found on the surface of infected cells. Excreted virions were devoid of gH and were not infectious. Virions excreted at the permissive temperature were infectious and contained gH and no loss of gH resulted from incubation of these virions at the non-permissive temperature. The temperaturesensitive phenotype apparently results from the loss of gH from virions during their transport to the cell surface, and since loss of gH is accompanied by loss of infectivity we conclude that gH is an essential component of the infectious virion.

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Importance of the Herpes Simplex VirusUL24Gene for Productive Ganglionic Infection in Mice

et al. 1998

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The UL24 gene of herpes simplex virus overlaps the viral thymidine kinase (tk) gene. Most previous studies of UL24 have examined UL24 mutants that have also contained tk and sometimes other mutations. To address the importance of UL24 for viral replication in cell culture and in infections of a mammalian host, we constructed a mutant virus containing a UL24 nonsense mutation that does not affect TK activity and a second mutant that contains clustered point mutations in UL24 and a mutation in tk that does not by itself affect the ability of the virus to replicate acutely in mouse ganglia or to reactivate from latent infection following corneal inoculation of mice. Both mutant viruses replicated in cells in culture and in the mouse eye, albeit less efficiently than wild type or control viruses. Both mutants were much more severely impaired for acute replication in trigeminal ganglia and for reactivation from latency following explant of these ganglia. Viral DNA and latency-associated transcripts were present, albeit at lower levels in ganglia infected with the nonsense mutant. These results indicate that UL24 is especially important for productive infection of mouse sensory ganglia and may have implications for the behaviors of certain tk mutants in pathogenesis.

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Genetic analysis of temperature-sensitive mutants of HSV-1: The combined use of complementation and physical mapping for cistron assignment

Cited by 99 publications

References 34 publications

DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus

DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus

Excretion of Non-infectious Virus Particles Lacking Glycoprotein H by a Temperature-sensitive Mutant of Herpes Simplex Virus Type 1: Evidence that gH Is Essential for Virion Infectivity

Importance of the Herpes Simplex VirusUL24Gene for Productive Ganglionic Infection in Mice

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