SUPPLEMENTARY DATA
Generation of null mutants of sar1, CG3542 and CG3542.
We obtained two deletions of sar1 by the imprecise excision of EP3575, which
remove 2114 and 1910 nucleotides (Fig. 4). Both deletions affect four exons of sar1,
which includes the first coding exon shared by all transcripts derived from the sar1
locus. The deletions remove the sequence encoding the N-terminus of the protein,
including the beta-strands thought to promote the GTP-dependent interaction with
lipid membranes (BI et al. 2002), and as such probably represent null alleles. Both
deletions are lethal when homozygous with a lethal period similar to the EP3575
stock. These deletions also fail to complement EP3575, suggesting that the original
EP insertion represents a loss of function allele of sar1.
The mobilisation of EP0719, which is inserted into the coding region of CG3542,
resulted in 6 lethal imprecise excisions. The putative protein encoded by this gene
shares 39% identity and 53% similarity with human Formin Binding Protein 11. The
deletions removed large parts of the coding region of CG3542 (Fig. 4), and embryos
homozygous for the deletions had the same lethal period as those homozygous for the
original insertion, consistent with the idea that EP0719 disrupts the function of
CG3542.
When we excised EP0863, which is inserted into the second exon of CG6386 we
recovered two imprecise excisions that removed 1258 and 1784 nucleotides (Fig. 4).
This gene is predicted to encode a kinase with a high degree of similarity to Vaccinia
Related Kinase 1 (VRK1). Both excisions delete most of the coding region of
1
CG6386, and are likely to represent null alleles of the gene (see Fig. 4). Embryos
homozygous for these deletions die at late embryonic or early larval stages, like those
homozygous for the original EP insertion.
REFERENCE
BI, X., R. CORPINA and J. GOLDBERG, 2002 Structure of the Sec23/24-Sar1 prebudding complex of the COPII vesicle coat. Nature 419 (6904): 271-277.
SUPPLEMENTARY TABLE
Breakpoints of deletions created by P-element excisions (cf. Figure 4).
The sequences represent the twenty first (left breakpoint) and twenty last (right
breakpoint) of the fragment that has been deleted. The sequence is given in the same
orientation as the coding strand of the gene.
Gene
Allele Left breakpoint
CG3542
#3.3
#9.4
#35.1
#18.2
#9.3
#15.5
#71
#28
#43
#53
sar1
CG6385
GAGACCTGCT
GAGTACGTGG
ACGTGAGCGG
CATCACTCGC
CATCACTCGC
CATCACTCGC
TTATGATTAA
TTATGATTAA
TATACTTGGC
TATACTTGGC
GGGAGCCGCC
ATATGAAGGC
GAAAAGAAGC
GATCACGAAA
GATCACGAAA
GATCACGAAA
AAAAAACTGA
AAAAAACTGA
CGCTATTTAA
CGCTATTTAA
Right breakpoint
.......
.......
.......
.......
.......
.......
.......
.......
.......
.......
2
CTTTGTTAAG
TGTTAAGGAA
CAGCGAGCAG
TGGATCGCGA
CCTCGCATTT
ACTCACCCGC
CACTACATTC
ACCCATACAG
TCACTGGACG
CCAGTTCGAA
GAAAGCGAAG
AGCGAAGATG
CTCGACGAGT
AACGAGGAGC
AAATTGAATG
AAAGAAAAAG
GACTTGGGTG
CCAGTGCGTC
AAAAAATCTC
GGAGGAGGCC