The T-DNA insertion in hcf155 reduces the
expression of a gene [gene designation VIPP1
(AAC27135.1)] that encodes a chloroplast protein that is located in
both the inner envelope and the thylakoid membrane. (a)
Localization of the T-DNA insertion site and identification of the
affected gene by Northern blot analysis. The hybridization probes
specific for the AAC27134 and AAC27135 genes that flank the T-DNA
insertion were generated by PCR. RNA loadings were 10 μg of total RNA
(WT and hcf155) and a 50% dilution in the case of
wild-type RNA. (b) Amino acid alignment of the
VIPP1/IM30 precursor proteins of Arabidopsis thaliana
(At) and Pisum sativum (Ps). The putative cleavage sites
are indicated. (c) Localization of the VIPP1 protein in
plastid fractions of A. thaliana wild-type plants with
the use of marker proteins of established topology: Tic40, subunit of
the protein translocation machinery of the inner envelope (IE) (29);
Toc160, subunit of the protein translocation machinery of the outer
envelope (OE) (16); RbcL, large subunit of the stromal (STR) enzyme
ribulose-1,5-bisphosphate carboxylase/oxygenase; LHCP, major
chlorophyll a/b-binding antenna protein
of photosystem II, a thylakoidal (THY) protein complex. Equal amounts
of protein (8 μg per lane) were loaded onto the gel.