Leucine at position 278 of the AIK-C measles virus vaccine strain fusion protein is responsible for reduced syncytium formation.
The live measles virus (MV) vaccine strain AIK-C was attenuated from the wild-type strain Edmonston by plaque purification at 33 degrees C. Strain AIK-C grew well at 33 degrees C with a mixture of small-and medium-sized plaques in Vero cells, but did not grow well at 40 degrees C. To investigate fusion inducibility, expression plasmids for the fusion (F) and haemagglutinin (H) protein regions of MV strains AIK-C (pAIK-F01 and pAIK-H) and Edmonston (pEdm-F and pEdm-H) were constructed. pEdm-F induced extensive cell fusion in B95a and Vero cells under the control of T7 RNA polymerase, whereas a sharp reduction in syncytium formation was observed when pAIK-F01 was used. Six amino acid differences were determined between pAIK-F01 and pEdm-F. Direct sequencing showed that the seed strain AIK-C contained either Leu or Phe at position 278 of the F protein. Experiments using recombinant F protein plasmids demonstrated that those with Leu at position 278 induced poor syncytium formation, while those with Phe at position 278 (Edmonston type) induced extensive cell fusion. Replacement of Phe with Leu at position 278 of pEdm-F reduced fusion-inducing capability. A full-length infectious clone of AIK-C with Leu at position 278 of the F protein was constructed. The rescued virus produced small plaques in Vero cells. However, the same rescued virus with Phe at position 278 produced large plaques. It was concluded that Leu at position 278 of the F protein of the MV vaccine strain AIK-C is responsible for the formation of small plaques.
Nakayama
Tetsuo
T
Department of Virology, Center for Basic Research1, and Division of Research and Development, Research Center for Biologicals2, The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8642, Japan.
Komase
Katsuhiro
K
Department of Virology, Center for Basic Research1, and Division of Research and Development, Research Center for Biologicals2, The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8642, Japan.
Uzuka
Rina
R
Department of Pediatrics, Tokyo Medical University, 6-7-1 Nishishinjyuku, Shinjyuku-ku, Tokyo 160-0023, Japan3.
Hoshi
Akiyoshi
A
Department of Pediatrics, Tokyo Medical University, 6-7-1 Nishishinjyuku, Shinjyuku-ku, Tokyo 160-0023, Japan3.
Okafuji
Takao
T
Department of Virology, Center for Basic Research1, and Division of Research and Development, Research Center for Biologicals2, The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8642, Japan.
D000818:Animals; D002522:Chlorocebus aethiops; D007930:Leucine; D008458:Measles Vaccine; D008459:Measles virus / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011817:Rabbits; D013329:Structure-Activity Relationship; D014613:Vaccines, Attenuated; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D008458:Measles Vaccine; D014613:Vaccines, Attenuated; D014760:Viral Fusion Proteins; D007930:Leucine