Antisense RNA to the first N-glycosylation gene, ALG7, inhibits protein N-glycosylation and secretion by Xenopus oocytes.
N-Glycosylation has been shown to affect the rate of glycoprotein transport through the secretory pathway. In order to identify the critical components in the N-glycosylation pathway that directly influence protein secretion, we have studied the effects of downregulation of the first gene in the dolichol pathway, ALG7, on the synthesis, glycosylation and secretion of native and heterologous proteins by Xenopus laevis oocytes. Our strategy involved the use of ALG7 antisense RNA (asRNA) to lower the effective abundance of the ALG7 protein in oocytes. The results showed that there was an inverse dose-response relationship between ALG7 asRNA and the amount of glycosylated and secreted proteins. These effects were also observed for heterologously expressed rat parotid amylase. Since ALG7 asRNA did not inhibit overall protein synthesis, we conclude that downregulation of ALG7 expression directly lowered protein export.
Kukuruzinska
M A
MA
Department of Oral Biology, Boston University Medical Center, Massachusetts 02118.
Apekin
V
V
Lamkin
M S
MS
Hiltz
A
A
Rodriguez
A
A
Lin
C C
CC
Paz
M A
MA
Oppenheim
F G
FG
D000681:Amylases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006031:Glycosylation; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D010306:Parotid Gland / Q000378:metabolism; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger; D051381:Rats; D012463:Saliva / Q000201:enzymology; D014982:Xenopus laevis
D016372:RNA, Antisense; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D000681:Amylases