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Journal of Virology, March 2000, p. 2383-2392, Vol. 74, No. 5
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
A Stable HeLa Cell Line That Inducibly Expresses
Poliovirus 2Apro: Effects on Cellular and Viral Gene
Expression
Angel
Barco,
Elena
Feduchi, and
Luis
Carrasco*
Centro de Biología Molecular
(CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain
Received 14 July 1999/Accepted 2 December 1999
A HeLa cell clone (2A7d) that inducibly expresses the gene for
poliovirus protease 2A (2Apro) under the control of
tetracycline has been obtained. Synthesis of 2Apro induces
severe morphological changes in 2A7d cells. One day after tetracycline
removal, cells round up and a few hours later die. Poliovirus
2Apro cleaves both forms of initiation factor eIF4G,
causing extensive inhibition of capped-mRNA translation a few hours
after protease induction.
Methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone, a selective
inhibitor of 2Apro, prevents both eIF4G cleavage and
inhibition of translation but not cellular death. Expression of
2Apro still allows both the replication of poliovirus and
the translation of mRNAs containing a picornavirus leader sequence,
while vaccinia virus replication is drastically inhibited. Translation
of transfected capped mRNA is blocked in 2A7d-On cells, while
luciferase synthesis from a mRNA bearing a picornavirus internal
ribosome entry site (IRES) sequence is enhanced by the presence of
2Apro. Moreover, synthesis of 2Apro in 2A7d
cells complements the translational defect of a poliovirus 2Apro-defective variant. These results show that poliovirus
2Apro expression mimics some phenotypical characteristics
of poliovirus-infected cells, such as cell rounding, inhibition of
protein synthesis and enhancement of IRES-driven translation. This cell
line constitutes a useful tool to further analyze 2Apro
functions, to complement poliovirus 2Apro mutants, and to
test antiviral compounds.
*
Corresponding author. Mailing address: Centro de
Biología Molecular (CSIC-UAM), Universidad
Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone:
34-91-397 8450. Fax: 34-91-397 4799. E-mail:
LCARRASCO{at}TRASTO.CBM.UAM.ES.

Present address: Howard Hughes Medical Institute, Center for
Neurobiology and Behavior, Columbia University, New York, NY
10032.
Journal of Virology, March 2000, p. 2383-2392, Vol. 74, No. 5
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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