Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - An avl9-1 apl2· vps1· ade2 ade3 strain cannot survive without a plasmid containing APL2 ADE3 and does not sector. The same strain background with a plasmid carrying the YLR114c gene can survive without the APL2 ADE3 plasmid and is white.
• Supplemental Figure 4a - (A) Alignment of two conserved regions between Avl9 proteins from human, A. mellifera (honey bee), C. elegans, and S. cerevisiae. Residues that are identical to the concensus are shaded red; similar residues are shaded yellow. The mutation in avl9-1 is indicated by an asterisk. Regions with similarity to Avl9 superfamily proteins are marked as AH1-5.
• Supplemental Figure 4b - (B) Multiple sequence alignment used in generating the tree shown in Figure 12. AH homology regions and uDENN, DENN regions are indicated. The uDENN region overlaps AH1 and a small part of AH2. Residues are colored according to chemical property.
• Supplemental Figure 4c - (C) Matrix showing percent identities and divergence between the AH1-4 and uDENN/DENN regions used for the phylogenetic analysis. Values were obtained by supplying the alignment shown in (B) to the MegAlign module from Lasergene 7 software. Divergence values are based on the rough phylogeny reconstructed by MegAlign and were not used for the tree shown in Figure 12. They are shown here because they are a better indication of distance between sequences than are percent identities.
• Supplemental Figure 3 - ER-to-Golgi transport is not defective in cells overexpressing Avl9p. Cells were grown as for Figure 10C, and CPY and Bgl2p were immuno-precipitated after 5 min pulse, 5 min chase.
• Supplemental Figure 2 - The temperature-sensitive rho3-1 allele is synthetically lethal in combination with vps1· apl2· at the normally permissive temperature of 24°C. In a cross, the triple mutant does not germinate or forms micro-colonies.