Characterization of LEF4 Ligand Binding Property and Its Role As Part of Baculoviral Transcription Machinery

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 2009 Jul 28

PMID 19633819

Authors

Sheeba Rasheedi

Madhuri Suragani

Soghra K Haq

Sudip Ghosh

Nasreen Z Ehtesham

Seyed E Hasnain

Affiliations

Soon will be listed here.

Abstract

Late expression factor 4 (LEF4) is one of the four identified subunits of Autographa californica nucleopolyhedrosis virus (AcNPV) encoded RNA polymerase that carries out transcription from viral late and very late promoters. This 464-amino acid baculovirus-encoded protein also harbors 5' mRNA capping activity that includes RNA 5' triphosphatase, nucleoside triphosphatase, and guanylyltransferase activities. Hydrolysis of 5' triphosphate RNA and free NTPs is metal ion dependent property of the protein. In the present communication, we describe the structural changes in the recombinant LEF4 protein following ligand binding. Metal ion binding causes some alteration in the conformation around aromatic amino acids whereas there is no effect on tryptophan fluorescence on GTP binding in absence and presence of metal ion. It is found that GTP and divalent cation cofactor produce some prominent changes in the secondary structure of the protein. Electrophoretic mobility shift assay (EMSA) shows that LEF4 is the probable factor that acts as anchor to dock the viral RNA polymerase on the very late polyhedrin promoter (Ppolh) facilitated by other factors.

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