Conformational aberrance of the sendai virus F0 protein in thapsigargin-treated cells allowing exit from the endoplasmic reticulum but causing arrest at the Golgi complex.
Thapsigargin and ionomycin inhibited the intracellular transport of the Sendai virus F0 protein. Depletion of Ca2+ from intracellular Ca2+ store(s) is critical for the inhibition, since ionomycin was more effective in the absence of extracellular Ca2+ than in its presence. Transport of F0 was arrested between the trans-Golgi complex and the plasma membrane [Ono, A. and Kawakita, M. (1994) J. Biochem. 116, 649-656], but only when thapsigargin was added before the synthesis of F0. This implies that F0 was committed to later arrest in the endoplasmic reticulum. Non-reducing SDS-polyacrylamide gel electrophoresis revealed a conformational abnormality of F0 immediately after pulse-labeling in thapsigargin-treated cells. The abnormality did not affect the exit of F0 from the endoplasmic reticulum, but paralleled its later arrest at the trans-Golgi stage. Pulse-labeled and 1-h-chased F0 was endoglycosidase H-resistant even in thapsigargin-treated cells, but was not recognized by mAb f-49, a monoclonal antibody that recognizes the corresponding F0 intermediate in uninhibited cells. The misfolded F0 may escape from recognition by means of the quality control system of the endoplasmic reticulum, but another system in the Golgi complex may complement the former. The arrested F0 was rapidly degraded.
Ono
A
A
Department of Physiological Chemistry, Tokyo Metropolitan Institute of Medical Science, Rinshoken.
Kawakita
M
M
D016475:3T3 Cells; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D011487:Protein Conformation / Q000187:drug effects*; D011498:Protein Precursors / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D013729:Terpenes / Q000494:pharmacology*; D019284:Thapsigargin; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D004791:Enzyme Inhibitors; C099057:F0 protein, Sendai virus; D011498:Protein Precursors; D013729:Terpenes; D014759:Viral Envelope Proteins; D015759:Ionomycin; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium