PMID- 10993946
OWN - NLM
STAT- MEDLINE
DCOM- 20001115
LR  - 20200213
IS  - 0022-1317 (Print)
IS  - 0022-1317 (Linking)
VI  - 81
IP  - Pt 10
DP  - 2000 Oct
TI  - Expression of unglycosylated mutated prion protein facilitates PrP(Sc) formation 
      in neuroblastoma cells infected with different prion strains.
PG  - 2555-2563
LID - 10.1099/0022-1317-81-10-2555 [doi]
AB  - Prion replication involves conversion of the normal, host-encoded prion protein 
      PrP(C), which is a sialoglycoprotein bound to the plasma membrane by a 
      glycophosphatidylinositol anchor, into a pathogenic isoform, PrP(Sc). In earlier 
      studies, tunicamycin prevented glycosylation of PrP(C) in scrapie-infected mouse 
      neuroblastoma (ScN2a) cells but it was still expressed on the cell surface and 
      converted into PrP(Sc); mutation of PrP(C) at glycosylation consensus sites 
      (T182A, T198A) produced low steady-state levels of PrP that were insufficient to 
      propagate prions in transgenic mice. By mutating asparagines to glutamines at the 
      consensus sites, we obtained expression of unglycosylated, epitope-tagged 
      MHM2PrP(N180Q,N196Q), which was converted into PrP(Sc) in ScN2a cells. Cultures 
      of uninfected neuroblastoma (N2a) cells transiently expressing mutated PrP were 
      exposed to brain homogenates prepared from mice infected with the RML, Me7 or 
      301V prion strains. In each case, mutated PrP was converted into PrP(Sc) as 
      judged by Western blotting. These findings raise the possibility that the N2a 
      cell line can support replication of different strains of prions.
FAU - Korth, Carsten
AU  - Korth C
AD  - Institute for Neurodegenerative Diseases1 and Departments of Neurology2 and 
      Biochemistry and Biophysics3, Box 0518, University of California, San Francisco, 
      CA 94143-0518, USA.
FAU - Kaneko, Kiyotoshi
AU  - Kaneko K
AD  - Institute for Neurodegenerative Diseases1 and Departments of Neurology2 and 
      Biochemistry and Biophysics3, Box 0518, University of California, San Francisco, 
      CA 94143-0518, USA.
FAU - Prusiner, Stanley B
AU  - Prusiner SB
AD  - Institute for Neurodegenerative Diseases1 and Departments of Neurology2 and 
      Biochemistry and Biophysics3, Box 0518, University of California, San Francisco, 
      CA 94143-0518, USA.
LA  - eng
GR  - AG02132/AG/NIA NIH HHS/United States
GR  - AG10770/AG/NIA NIH HHS/United States
GR  - NS14069/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (PrPSc Proteins)
RN  - 0 (Prions)
SB  - IM
MH  - Amino Acid Substitution
MH  - Animals
MH  - Consensus Sequence
MH  - Glycosylation
MH  - Mice
MH  - Mutagenesis, Site-Directed
MH  - Neuroblastoma/metabolism/*virology
MH  - PrPSc Proteins/*biosynthesis/genetics
MH  - Prions/*biosynthesis/genetics/*physiology
MH  - Quantitative Structure-Activity Relationship
MH  - Transfection
MH  - Tumor Cells, Cultured
MH  - *Virus Replication
EDAT- 2000/09/20 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/09/20 11:00
PHST- 2000/09/20 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/09/20 11:00 [entrez]
AID - 10.1099/0022-1317-81-10-2555 [doi]
PST - ppublish
SO  - J Gen Virol. 2000 Oct;81(Pt 10):2555-2563. doi: 10.1099/0022-1317-81-10-2555.