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. 2014 Feb;88(3):1461–1472. doi: 10.1128/JVI.02384-13

FIG 7.

FIG 7

The VSV P protein contains a phosphotyrosine residue (A) BHK 21 cells either uninfected or infected with VSV (MOI, 1.0) were harvested at 16 h of postinfection. Uninfected cell extract (lanes 1, 4, and 7) and infected-cell extracts either untreated (lanes 2, 5, and 8) or treated with calf intestine alkaline phosphatase (CIP) (lanes 3, 6, and 9) were resolved by 10% SDS-PAGE and blotted to a PVDF membrane. The same membrane was then probed with phosphotyrosine antibody (lanes 1 to 3), stripped, and probed with polyclonal anti-P antibody (lanes 4 to 6) and then polyclonal anti-N antibody (lanes 7 to 9). The values to the left are molecular sizes in kilodaltons. (B) Phosphoprotein sequences from 27 different strains of 11 different vesiculoviruses (listed in the ViPR database) were aligned by using MUSCLE software. The tyrosine residue, indicated by the arrow, is conserved across all of them. The sequences aligned are those of Chandipura virus (CHPV1 and CHPV2; UniProt Knowledgebase [UniProtKB] accession no. of P proteins, E3T3B5 and E3T3A5, respectively; strains CIN 0327 and CIN 0451), Cocal virus (COCV; UniProtKB accession no. of P protein, B3FRK7; strain Indiana 2), Jurona virus (JURV; UniProtKB accession no. of P protein, I1SV83; strain unknown [accession no. HM566194]), Maraba virus (MARAV; UniProtKB accession no. of P protein, F8SPF1; strain unknown [accession no. HQ660076]), Perinet virus (PERV; UniProtKB accession no. of P protein, I1SV88; strain unknown [accession no. HM566195]), Pike fry rhabdovirus (PFRV; UniProtKB accession no. of P protein, C3VM12; strain F4), Spring viremia of carp virus (SVCV-1-5; UniProtKB accession no. of P protein, A8VJA2, Q91DS2, Q91DS5, A0FCR4, and Q1KK77; strain BJ0505-2), Fijan reference strains ATCC VR-1390 and ATCC VR-1390 (A1 and A2), Vesicular stomatitis Alagoas virus (VSAV; UniProtKB accession no. of P protein, B3FRL2; strain Indiana 3), VSV strain Indiana (VSIV-1 to -4; UniProtKB accession no. of P proteins, Q8B0I3, Q8B0H3, P03520], and Q8B0H8; strains 98COE, 94GUB, unknown [accession no. J02428], and 85CLB), Vesicular stomatitis New Jersey virus (VSNJV-1 to -9; UniProtKB accession no. of P proteins, I7CGJ1, I7CGJ9, I7CGL3, I7CGM8, I7CGK3, I7CGL8, I7CGJ5, I7CGL3, and I7CGM3; strains NJ0783NCP, NJ92CRB, NJ1184HDB, NJ95COB, NJ0703CRB6, NJ89GAS, NJ0185PNB, NJ92CLB, and NJ0405NME), Yug Bogdanovac virus (YBV; UniProtKB accession no. of P protein, K4FFQ0; strain unknown [accession no. JF911700]).