Isolation of a peptide inhibitor of human rhinovirus.
Cell culture-based transdominant genetic techniques provide new methods for discovering peptide/RNA modulators of cellular pathways. We applied this technology to isolate a peptide inhibitor of human rhinovirus. A green fluorescent protein (GFP)-scaffolded library of cDNA fragments was expressed in HeLa cells from a retroviral vector and screened for inhibitors of rhinovirus-mediated cell killing. A DNA clone, I421, increased cell survival in an HRV14 challenge assay from less than 0.5% to greater than 60%. It encodes a 53-amino-acid C-terminal extension of the GFP scaffold. Particular subclones of Hela cells expressing I421 (exemplified by I421dp3) show a delay in virus production and a 50-fold decrease in viral RNA levels at 6-8 h postinfection. HRV2, HRV14, and HRV16 show a dramatic decrease in plaque-forming ability on I421dp3 while Coxsackievirus B3 showed a small reduction. Levels of ICAM-1, the receptor for the main rhinovirus serotype, are not altered in I421dp3.
Poritz
Mark A
MA
Deltagen Proteomics, Inc., 615 Arapeen Drive, Suite 300, Salt Lake City, UT 84108, USA. maporitz83@post.harvard.edu
Malmstrom
Sharon
S
Schmitt
Amy
A
Kim
Marianne K-H
MK
Zharkikh
Ludmilla
L
Kamb
Alexander
A
Teng
David H-F
DH
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods; D015723:Gene Library; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D010920:Placenta / Q000737:chemistry; D012190:Retroviridae / Q000235:genetics; D012229:Rhinovirus / Q000187:drug effects* / Q000502:physiology; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D018076:DNA, Complementary; D010455:Peptides