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THE PLANT CELL, Vol 7, Issue 5 589-598, Copyright © 1995 by American Society of Plant Biologists
The Activation of the Potato PR-10a Gene Requires the Phosphorylation of the Nuclear Factor PBF-1
C. Despres, R. Subramaniam, D. P. Matton and N. Brisson
Department of Biochemistry, Universite de Montreal, Montreal, Canada H3C 3J7
The pathogenesis-related gene PR-10a (formerly STH[middot]2) is induced in
various organs of potato after wounding, elicitor treatment, or infection
by Phytophthora infestans. Deletion analysis of the promoter of the PR-10a
gene enabled us to identify a 50-bp region, located between positions -155
and -105, necessary for the elicitor responsiveness of the
[beta]-glucuronidase reporter gene in transgenic potato plants. Within this
region, a 30-bp sequence, located between positions -135 and -105, was
necessary for the activation of the promoter by the elicitor. However,
strong promoter activity after elicitor treatment required the presence of
a 20-bp sequence located between positions -155 and -135. The region
between -135 and -105 was specifically recognized by two nuclear factors,
PBF-1 (PR-10a Binding Factor 1) and PBF-2, and binding of PBF-1 was
coordinated with the accumulation of the PR-10a mRNA. Gel shift assays
using nuclear extracts pretreated with sodium deoxycholate or alkaline
phosphatase suggested that PBF-1 is a multimeric factor in which at least
one of the constituent proteins can be phosphorylated. Treatment with
alkaline phosphatase also indicated that binding of PBF-1 is positively
regulated by phosphorylation and that it is phosphorylated only in tissues
in which PR-10a is expressed. The use of protein phosphatase and kinase
inhibitors in vivo provided additional evidence that wounding and elicitor
treatment induce the phosphorylation of PBF-1 and that this phosphorylation
is associated with gene activation.
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