JBC Avanti Polar Lipids

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Volume 270, Number 17, Issue of April 28, pp. 10105-10112, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Poliovirus Protein 2C Contains Two Regions Involved in RNA Binding Activity

Pedro L. Rodrguez , Luis Carrasco

Poliovirus protein 2C is involved in poliovirus RNA replication, although the exact function of 2C is still unknown. Recently, it was shown that 2C can be purified to high levels when expressed as a fusion protein with maltose-binding protein (MBP). Evidence was presented that 2C has ATPase and GTPase activities; preliminary results also indicated that 2C interacts with RNA (Rodrguez, P. L., and Carrasco, L. (1993) J. Biol. Chem. 268, 8105-8110). In the present study, 20 variants of 2C have been generated, and their NTPase and RNA binding activities were analyzed. Moreover, an easy procedure to obtain genuine 2C after factor Xa cleavage of an MBP-2C fusion protein is described. This work has determined that 2C has two regions involved in RNA binding: a NH-terminal region located between amino acids 21 and 45 and a COOH-terminal region involving an Arg-rich region located between amino acids 312 and 319. Deletion of either the NH- or COOH-terminal RNA-binding region abolishes RNA binding. Deletion of an internal region of protein 2C that includes the nucleotide-binding motif does not affect RNA binding, whereas this deletion destroys ATPase and GTPase activities. Therefore, the NTPase activity and the RNA binding capacity of protein 2C are located in different regions of the molecule.



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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.