Fig.1Technical route of fusion PCR (Appendix 1)
Fig. 1 Experimental procedure of the construction of the recombinant strains (Appendix 2)
R and H differently mean left and right homologous arm; F means FLAG tag; K means gene KANMX6.
Appendix 3:Identification ofthe RAVEcomplex by MALDI-TOF mass spectrometry
Fig. 1PMF ofRav1p a: Mass spectrum; b: Result of Mascot
Tab. 1Second-order mass spectrometry matching peptide of Rav1p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1195.6197 / 1194.612 / 1194.603 / 47 / 0.48 / R.FISNDFTVPR.W
1448.7844 / 1447.777 / 1447.765 / 63 / 0.013 / K.FCVIIDNWIIR.Q
1496.8701 / 1495.863 / 1495.851 / 22 / 1.20E+02 / K.LSFLVHDLQGVIR.H
1799.8954 / 1798.888 / 1798.874 / 78 / 0.00032 / K.SLDLTDPFTYQSIGSR.K
2063.1143 / 2062.107 / 2062.089 / 13 / 7.20E+02 / R.SFLVEDPALLYLYNHLR.N
Note: We got the parameters from the result of Mascot and summarized as table 1.
1111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111
Fig. 2 PMF of Leu1p a: Mass spectrum; b: Result of Mascot
Tab. 2Second-order mass spectrometry matching peptide of Leu1p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1282.609 / 1281.601 / 1281.6142 / 24 / 13 / K.IFQEAGFEWR.E
1490.665 / 1489.658 / 1489.666 / 52 / 0.021 / K.QFGVPYFGMSDAR.Q + Oxidation (M)
2224.041 / 2223.033 / 2223.0596 / 33 / 1.1 / K.QAEAEGLDKIFQEAGFEWR.E
2343.187 / 2342.179 / 2342.1907 / 37 / 0.82 / K.ILDSDGNVLVDHFEIEPFRK.H
Note: We got the parameters from the result of Mascot and summarized as table 2.
Fig. 3 PMF of Vma1p a: Mass spectrum; b: Result of Mascot
Tab. 3Second-order mass spectrometry matching peptide of Vma1p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1307.7153 / 1306.708 / 1306.699 / 77 / 0.00051 / K.VGHDNLVGEVIR.I
1333.7201 / 1332.713 / 1332.704 / 59 / 0.034 / K.LSADYPLLTGQR.V
1646.8004 / 1645.793 / 1645.778 / 46 / 0.43 / K.FYDSNYPEFPVLR.D
Note: We got the parameters from the result of Mascot and summarized as table 3.
Fig. 4 PMF of Vma2p a: Mass spectrum; b: Result of Mascot
Tab. 4Second-order mass spectrometry matching peptide of Vma2p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1244.684 / 1243.676 / 1243.667 / 49 / 0.3 / K.AVEQGFNVKPR.L
1472.762 / 1471.754 / 1471.742 / 52 / 0.16 / R.IFDGSGRPIDNGPK.V
1525.890 / 1524.883 / 1524.866 / 51 / 0.096 / K.GIYPPINVLPSLSR.L
1703.914 / 1702.906 / 1702.889 / 48 / 0.3 / R.YNEIVNLTLPDGTVR.Q
Note: We got the parameters from the result of Mascot and summarized as table 4.
Fig. 5 PMF of Rav2p a: Mass spectrum; b: Result of Mascot
Tab. 5Second-order mass spectrometry matching peptide of Rav2p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1699.7572 / 1698.75 / 1698.771 / 11 / 4.6 / R.GVTFDNRVVMECEK.L + Oxidation (M)
1766.8026 / 1765.795 / 1765.817 / 63 / 3.10E-05 / R.SFCDQIRNQVTNER.N
Note: We got the parameters from the result of Mascot and summarized as table 5.
Fig. 6PMF of Skp1p a: Mass spectrum; b: Result of Mascot
Tab.6Second-order mass spectrometry matching peptide of Skp1p
Observed / Mr(expt) / Mr(calc) / Score / Expect / Peptide1176.6177 / 1175.61 / 1175.584 / 53 / 0.15 / K.VIEWAEHHR.D
1445.7965 / 1444.789 / 1444.752 / 57 / 0.04 / M.VTSNVVLVSGEGER.F
Note: We got the parameters from the result of Mascot and summarized as table 6.
Appendix 4:Multiple alignment of the primary structure of the conserved region of Leu1p