title: A positively charged lysine residue at VP2 131 position allows for the enhanced adaptability of foot-and-mouth disease virus serotype A in BHK-21 cells.
abstract: Field outbreak strains of foot-and-mouth disease virus (FMDV) infect host cells through certain Arg-Gly-Asp (RGD) dependent integrin family of cellular receptors. In contrast, FMDV adapted in non-host cell cultures are reported to acquire the ability to infect cells via heparin sulphate (HS) or other unidentified cell surface molecules. It has been reported that during the serial passage of FMDV serotype A in BHK-21 cell culture, VP2 E131K (E2131K) substitution was fixed within the heparin sulphate binding site. The fixation of positively charged residue at position VP2 131 of serotype A is considered to associate with the ability to utilise alternative receptor. In this study, an infectious full-length cDNA clone for Indian FMDV vaccine strain A IND 40/2000 was constructed. Through site-directed mutagenesis on the cDNA clone, recombinant virus containing positive charged amino acid residue at position VP2 131 was rescued. The recombinant mutated virus was shown to have specific and strong affinity for HS and demonstrated an enhanced infectivity in BHK-21 cell line. The introduction of lysine residue at VP2 131 position that allows cell culture adaptation of FMDV serotype A could be exploited for the generation of vaccine seed stocks with improved growth properties in BHK-21 cell line.
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India.
authors.forename: Jitendra K
authors.initials: JK
authors.lastname: Biswal
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India.
authors.forename: Jajati K
authors.initials: JK
authors.lastname: Mohapatra
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India.
authors.forename: Punam
authors.initials: P
authors.lastname: Bisht
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India.
authors.forename: Saravanan
authors.initials: S
authors.lastname: Subramaniam
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India.
authors.forename: Aniket
authors.initials: A
authors.lastname: Sanyal
authors.affiliation: Project Directorate on Foot-and-Mouth Disease (ICAR), Mukteswar, 263138 Nainital, Uttarakhand, India. Electronic address: pattnaikb@gmail.com.
authors.forename: Bramhadev
authors.initials: B
authors.lastname: Pattnaik
mesh_terms: D000818:Animals; D001483:Base Sequence; D036022:Capsid Proteins / Q000737:chemistry / Q000502:physiology*; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000502:physiology*; D008239:Lysine / Q000378:metabolism*; D011995:Recombination, Genetic; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; C442459:VP2 protein, Foot-and-mouth disease virus; D008239:Lysine
keywords: Cell receptor; FMDV serotype A; Heparan sulphate; Infectious cDNA clone; Vaccine strain