Functional characterization of Ebola virus L-domains using VSV recombinants.
VSV recombinants containing the overlapping L-domain sequences from Ebola virus VP40 (PTAPPEY) were recovered by reverse-genetics. Replication kinetics of M40-WT, M40-P24L, and M40-Y30A were indistinguishable from VSV-WT in BHK-21 cells, whereas the double mutant (M40-P2728A) was defective in budding. Insertion of the Ebola L-domain region into VSV M protein was sufficient to alter the dependence on host proteins for efficient budding. Indeed, M40 recombinants containing a functional PTAP motif specifically incorporated endogenous tsg101 into budding virions and were dependent on tsg101 expression for efficient budding. Thus, VSV represents an excellent negative-sense RNA virus model for elucidating the functional aspects and diverse host interactions associated with the L-domains of Ebola virus.
Irie
Takashi
T
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3800 Spruce Street, Philadelphia, PA 19104-6049, USA.
Licata
Jillian M
JM
Harty
Ronald N
RN
D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism; D029043:Ebolavirus / Q000502:physiology*; D056827:Endosomal Sorting Complexes Required for Transport; D005816:Genetic Complementation Test / Q000379:methods*; D019142:Hemorrhagic Fever, Ebola / Q000821:virology; D006801:Humans; D009698:Nucleoproteins / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology*; D011995:Recombination, Genetic; D014157:Transcription Factors / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism; D014758:Viral Core Proteins / Q000502:physiology*; D014779:Virus Replication; D000099177:Tsg101 Protein
D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D009698:Nucleoproteins; D014157:Transcription Factors; D014758:Viral Core Proteins; D000099177:Tsg101 Protein; C084819:nucleoprotein VP40, Ebola virus