Table 2. Affinity of substrate RNAs binding to H/ACA RNPs.
| Guide RNA | Substrate RNA | Proteins | K D, nM |
|---|---|---|---|
| snR34 | 5′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 53 ± 22 |
| snR34 | 3′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 100 ± 30 |
| snR5 | 5′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 160 ± 50 |
| snR5 | 3′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 330 ± 70 |
| snR34 | 5′ substrate | Cbf5p–Nop10p–Gar1p | 50 ± 20 |
| snR34 | 3′ substrate | Cbf5p–Nop10p–Gar1p | 90 ± 20 |
| snR34 Δ H box | 3′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 90 ± 30 |
| snR34 Δ ACA box | 3′ substrate | Cbf5p–Nop10p–Gar1p–Nhp2p | 150 ± 30 |
The dissociation constants (KD) of substrate RNA binding to reconstituted H/ACA RNPs was determined by nitrocellulose filtration (Figure 4).