TABLE 1.
Genotypes and phenotypes of relevant CLN2 mutants and CLN2 domain fusions
| Genotype | Name | No. of sites mutated | Half-life (min)a | Binding
|
|
|---|---|---|---|---|---|
| Grr1 | Cdc28 | ||||
| Cln2 mutants | |||||
| CLN2 | Wild type | 0 | 8 | ++ | + |
| CLN2(S518A) | 1 | 6 | NDc | + | |
| CLN2(S396,427A) | D35 | 2 | 35 | ND | + |
| CLN2(Δ376-514) | ΔHN | 5b | 55 | ND | + |
| CLN2(Δ376-545) | ΔC | 6b | 60 | ND | + |
| CLN2(T311,381,405,430A S396,427,518A) | 4T3S | 7 | 60 | − | + |
| CLN2(S396,427A T405,430A) | M46 | 4 | 55 | − | + |
| CLN2(S396,427D T405,430D) | M4D | 4 | 55 | ND | ND |
| cln2Δxs | Cln2Δxs | 0 | 30 | ND | − |
| cln2Δxs(T311,381,405,430A S396,427,518A) | ClnΔxs(4T3S) | 0 | 30 | ND | − |
| GST-Cln2 fusions | |||||
| GST | GST | na | 166 | − | − |
| GST-CLN2C(376-545) | GST-2C(wt) | 0 | 41 | − | − |
| GST-CLN2(376-545; T405,430A S396,427A) | GST-2C(M46) | 6 | 38 | − | − |
| GST-CLN2(376-431) | GST-PD | 0 | 93 | − | − |
| GST-CLN2(396-431) | GST-D | 0 | 134 | − | − |
| Cln2-ΔN-Sic1 fusions | |||||
| CLN2C(376-545)-ΔN105-SIC1 | 2C(wt)-ΔN-Sic1 | 0 | 18 | ++ | + |
| CLN2C(376-545; T405,430A S396,427A)-ΔN105-SIC1 | 2C(M46)-ΔN-Sic1 | 4 | ∼180 | − | + |
| CLN2C(396-431)-ΔN105-SIC1 | D-Sic1 | 0 | 150 | + | + |
| CLN2C(376-403)-ΔN105-SIC1 | P-Sic1 | 0 | 90 | − | + |
| CLN2C(396-431)-ΔN105-SIC1 | PD-Sic1 | 0 | 41 | ++ | + |
a
Cln2 half-life was determined by densitometric analysis.
b
Number of sites deleted.
c
ND, not determined.