Yeast TGN resident proteins that frequently cycle between the TGN and endosomes are much more slowly transported to the prevacuolar/late endosomal compartment (PVC) than other proteins. However, TGN protein transport to the PVC is accelerated in mutants lacking function of Inp53p. Inp53p contains a SacI polyphosphoinositide phosphatase domain, a 5-phosphatase domain, and a proline-rich domain. Here we show that all three domains are required to mediate "slow delivery" of TGN proteins into the PVC. Although deletion of the proline-rich domain did not affect general membrane association, it caused localization to become less specific. The proline-rich domain was shown to bind to two proteins, including clathrin heavy chain, Chc1p. Unlike chc1 mutants, inp53 mutants do not mislocalize TGN proteins to the cell surface, consistent with the idea that Chc1p and Inp53p act at a common vesicular trafficking step but that Chc1p is used at other steps also. Like mutations in the AP-1 adaptor complex, mutations in INP53 exhibit synthetic growth and transport defects when combined with mutations in the GGA proteins. Taken together with other recent studies, our results suggest that Inp53p and AP-1/clathrin act together in a TGN-to-early endosome pathway distinct from the direct TGN-to-PVC pathway mediated by GGA/clathrin.
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| Gene | Phenotype | Experiment Type | Mutant Information | Strain Background | Chemical | Details |
|---|---|---|---|---|---|---|
| INP53 | protein secretion: abnormal Reporter: pro-alpha-factor | classical genetics | reduction of function Allele: inp53-sac1 C421A, C424A, R427A; Sac1 domain mutated | Other | Details: due to Kex2p depletion | |
| INP53 | protein secretion: abnormal Reporter: pro-alpha-factor | classical genetics | reduction of function Allele: inp53-5ptase1 D746A, D748A; 5-phosphatase domain mutated | Other | Details: due to Kex2p depletion | |
| INP53 | protein secretion: abnormal Reporter: pro-alpha-factor | classical genetics | reduction of function Allele: inp53-ΔC C-terminal truncation 910-end; proline-rich domain deleted | Other | Details: due to Kex2p depletion | |
| INP53 | protein transport: increased rate Reporter: A(F-A)-ALP cytoplasmic domain of Ste13p (F85A, F87A) fused to transmembrane and lumenal domains of Pho8p | classical genetics | reduction of function Allele: inp53-sac1 C421A, C424A, R427A; Sac1 domain mutated | Other | Details: slow delivery rate from TGN to PVC is accelerated | |
| INP53 | protein transport: increased rate Reporter: A(F-A)-ALP cytoplasmic domain of Ste13p (F85A, F87A) fused to transmembrane and lumenal domains of Pho8p | classical genetics | reduction of function Allele: inp53-5ptase1 D746A, D748A; 5-phosphatase domain mutated | Other | Details: slow delivery rate from TGN to PVC is accelerated | |
| INP53 | protein transport: increased rate Reporter: A(F-A)-ALP cytoplasmic domain of Ste13p (F85A, F87A) fused to transmembrane and lumenal domains of Pho8p | classical genetics | reduction of function Allele: inp53-ΔC C-terminal truncation 910-end; proline-rich domain deleted | Other | Details: slow delivery rate from TGN to PVC is accelerated | |
| INP53 | protein/peptide accumulation: decreased Reporter: Kex2p | classical genetics | reduction of function Allele: inp53-sac1 C421A, C424A, R427A; Sac1 domain mutated | Other | Details: accelerated Kex2p turnover | |
| INP53 | protein/peptide accumulation: decreased Reporter: Kex2p | classical genetics | reduction of function Allele: inp53-5ptase1 D746A, D748A; 5-phosphatase domain mutated | Other | Details: accelerated Kex2p turnover | |
| INP53 | protein/peptide accumulation: decreased Reporter: Kex2p | classical genetics | reduction of function Allele: inp53-ΔC C-terminal truncation 910-end; proline-rich domain deleted | Other | Details: accelerated Kex2p turnover |
Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.
| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.