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A
ipmi lsu1-1
LB
-89 AACCCCTTTA GGTTTTTTCT AACACACGCC TCTAATCTCC GCGCACACAC ACACACCCTC ACGAAGAAGA AGAAGACGAA
-9 GAAGAATCAA TGGCTTCTGT TATCTCTTCC TCTCCTTTTC TATGCAAATC ATCCTCCAAG GTTATTCATC TATCCTTCAT
ipmi lsu1-2
-169
-89
-9
CTAGAGACTC TCTCACAGTC AACGCAGCTT ATGTGTCATA GTAAGACTTT TTGTCTACTA TAGTAGAAAG ACGAATTTAT
AACCCCTTTA GGTTTTTTCT AACACACGCC TCTAATCTCC GCGCACACAC ACACACCCTC ACGAAGAAGA AGAAGACGAA
GAAGAATCAA TGGCTTCTGT TATCTCTTCC TCTCCTTTTC TATGCAAATC ATCCTCCAAG GTTATTCATC TATCCTTCAT
ipmi lsu1-3
-89
-9
LB
LB
LB
LB
AACCCCTTTA GGTTTTTTCT AACACACGCC TCTAATCTCC GCGCACACAC ACACACCCTC ACGAAGAAGA AGAAGACGAA
GAAGAATCAA TGGCTTCTGT TATCTCTTCC TCTCCTTTTC TATGCAAATC ATCCTCCAAG GTTATTCATC TATCCTTCAT
Sequence of the 5’ upstream region, of exon 1 (bold letters) and a part of intron 1 (grey letters) of the IPMI LSU gene
(At4g13430). The 5’ terminal nucleotides of three different cDNA clones are given in blue (cDNA clones RAFL 05-19-F17 and
RAFL 09-81-N04) and red (RAFL 05-21-C12) letters. In ipmi lsu1-2 (SAIL 369G04) and ipmi lsu1-3 (GABI Kat 452D03) two left
borders (LB) were detected. In these lines the insertion deleted 32 (GABI Kat 452D03) and 44 (SAIL 369G04) nucleotides
(underlined) in the genomic sequence. In the line ipmi lsu1-1 (SALK 029510) only a single left border was found. Numbering is
given in respect the ATG. The underlined A is position +1.
Knill et al., Supplemental Figure 2A
B
Stop P S Y S L L I H V D F P D M K P F T I E Y I L Q E E S M A S V I S S S …
ipmi lsu1-3
Stop S P R T H T H P H E E E E D E E E S M A S V I S S S …
wt
target P
cp
mt
Predotar
else
cp
mt
else
ipmi lsu1-3
0.875 0.014 0.012
0.04 0.01
0.96
wt
0.973 0.023 0.021
0.96 0.02
0.04
N-terminal amino acid sequences from the IPMI LSU1 wild-type and the ipmi lsu1-3 alleles. In ipmi lsu1-3 the T-DNA
insertion upstream of the “normal” predicted ATG (red) extends the reading frame by introducing an in frame translation
start codon (blue). This reduces the probability for the import into chloroplasts (cp) predicted by target P
(http://www.cbs.dtu.dk/services/TargetP/) and Predotar (http://urgi.versailles.inra.fr/predotar/predotar.html). Mt:
mitochondria; else: elsewhere.
Knill et al., Supplemental Figure 2B