PMID- 2548078
OWN - NLM
STAT- MEDLINE
DCOM- 19890921
LR  - 20210526
IS  - 0270-7306 (Print)
IS  - 1098-5549 (Electronic)
IS  - 0270-7306 (Linking)
VI  - 9
IP  - 6
DP  - 1989 Jun
TI  - Use of a glucocorticoid-inducible promoter for expression of herpes simplex virus 
      type 1 glycoprotein gC1, a cytotoxic protein in mammalian cells.
PG  - 2303-14
AB  - Abundant expression of herpes simplex virus type 1 glycoprotein gC (gC1) in 
      transfected mammalian cells has not previously been achieved, possibly because 
      gC1 protein is toxic to cells. To approach this problem, the gC1 coding sequence 
      was placed under the control of the weak but inducible glucocorticoid-responsive 
      promoter from the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). As 
      controls to evaluate for gC1 cytotoxicity, the MMTV LTR promoter was used to 
      express glycoprotein gD1, and a strong, constitutive promoter from the Moloney 
      murine sarcoma virus LTR was used to express gC1. L cells were transfected with 
      these constructs, and a clone expressing gC1 from the inducible MMTV LTR promoter 
      was analyzed. In the absence of glucocorticoid (dexamethasone) stimulation, only 
      a low level of gC1 mRNA expression was detected; after overnight stimulation with 
      dexamethasone, transcription increased approximately 200-fold. Abundant gC1 
      protein that was functionally active in that it bound complement component C3b, 
      was produced. From passages 5 through 26 (70 cell population doublings), the 
      gC1-producing clone became less responsive to overnight dexamethasone 
      stimulation. The block to gC1 expression occurred at the level of transcription 
      and was associated with hypermethylation of the MMTV LTR DNA. Treatment of the 
      clone with 5-aza-2'-deoxycytidine partially reversed the block in gC1 protein 
      production. Late-passage cells assumed a gC1-negative phenotype that appeared to 
      offer a selective growth advantage, which suggested that gC1 was cytotoxic. 
      Several findings support this view: (i) some cells expressing gC1 after overnight 
      stimulation with dexamethasone assumed bizarre, syncytial shapes; (ii) continuous 
      stimulation with dexamethasone for 5 weeks resulted in death of most cells; (iii) 
      cells transfected with gC1 under the control of the strong Moloney murine sarcoma 
      virus promoter assumed bizarre shapes, and stable gC1-expressing clones could not 
      be established; and (iv) cells induced to express gD1 retained a normal 
      appearance after overnight stimulation or 15 weeks of continuous stimulation with 
      dexamethasone. The inducible MMTV LTR promoter is useful for expressing gC1 and 
      may have applications for expressing other cytotoxic proteins.
FAU - Friedman, H M
AU  - Friedman HM
AD  - Department of Medicine, University of Pennsylvania School of Medicine, 
      Philadelphia.
FAU - Yee, A
AU  - Yee A
FAU - Diggelmann, H
AU  - Diggelmann H
FAU - Hastings, J C
AU  - Hastings JC
FAU - Tal-Singer, R
AU  - Tal-Singer R
FAU - Seidel-Dugan, C A
AU  - Seidel-Dugan CA
FAU - Eisenberg, R J
AU  - Eisenberg RJ
FAU - Cohen, G H
AU  - Cohen GH
LA  - eng
GR  - AI-18289/AI/NIAID NIH HHS/United States
GR  - FO6 TWO1162/TW/FIC NIH HHS/United States
GR  - HL 28220/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (Cytotoxins)
RN  - 0 (DNA, Viral)
RN  - 0 (Glucocorticoids)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (glycoprotein D, Human herpesvirus 1)
RN  - 0 (glycoprotein gC, herpes simplex virus type 1)
RN  - 7S5I7G3JQL (Dexamethasone)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Cloning, Molecular
MH  - Cytotoxins/biosynthesis/*genetics
MH  - DNA, Viral/genetics
MH  - Dexamethasone/*pharmacology
MH  - *Genes, Viral
MH  - Glucocorticoids/*genetics
MH  - Immunoblotting
MH  - Mammary Tumor Virus, Mouse/genetics
MH  - Plasmids
MH  - Promoter Regions, Genetic/*drug effects
MH  - RNA, Messenger/biosynthesis/genetics
MH  - RNA, Viral/biosynthesis/genetics
MH  - Simplexvirus/drug effects/*genetics
MH  - Transfection
MH  - Viral Envelope Proteins/biosynthesis/*genetics
PMC - PMC362303
EDAT- 1989/06/01 00:00
MHDA- 1989/06/01 00:01
PMCR- 1989/06/01
CRDT- 1989/06/01 00:00
PHST- 1989/06/01 00:00 [pubmed]
PHST- 1989/06/01 00:01 [medline]
PHST- 1989/06/01 00:00 [entrez]
PHST- 1989/06/01 00:00 [pmc-release]
AID - 10.1128/mcb.9.6.2303-2314.1989 [doi]
PST - ppublish
SO  - Mol Cell Biol. 1989 Jun;9(6):2303-14. doi: 10.1128/mcb.9.6.2303-2314.1989.