TABLE 1.
Targeted residues and resultant mutant activities
| Residuea | Reason(s) for selectionb | Mutation(s) | HIV-1 infectivity (mean % ± SE)c |
|---|---|---|---|
| Arg-3 | i | R3A | 80.6 ± 16.2 |
| R3A/K14A/K16A | -4.3 ± 5.7 | ||
| Leu-10 | ii | L10A | 50.1 ± 3.9 |
| Ile-11 | ii | I11A | 68.9 ± 14.8 |
| L10A/I11A | 8.0 ± 2.3 | ||
| Lys-14 | i, ii | K14A | 57.9 ± 0.9 |
| K14E | 16.6 ± 1.6 | ||
| Met-15 | ii, iii | M15E | 34.7 ± 0.1 |
| Lys-16 | i | K16A | 91.3 ± 0.2 |
| K16E | 42.0 ± 15.5 | ||
| K14A/K16A | -3.6 ± 6.3 | ||
| Tyr-18 | ii, iii | Y18A | 96.6 ± 1.3 |
| M15E/Y18A | 28.0 ± 0.6 | ||
| Pro-19 | ii | P19A | 107 ± 13.5 |
| P19E | 20.0 ± 6.9 | ||
| His-20 | iv | H20A | 94.8 ± 13.3 |
| H20E | 80.8 ± 9.8 | ||
| Trp-21 | ii, iii | W21A | -4.8 ± 4.7 |
| W21E | -3.4 ± 3.3 | ||
| M15E/Y18A/W21A | -3.1 ± 6.7 | ||
| Val-28 | ii | V28E | 103 ± 12.0 |
| Leu-40 | v | L40E | 42.9 ± 12.1 |
| Pro-41 | v | P41E | 89.0 ± 7.2 |
| L40E/P41E | 6.4 ± 0.3 | ||
| Ile-42 | ii | I42A | 19.4 ± 1.7 |
| Phe-43 | ii, iii | F43A | 41.3 ± 5.4 |
| I42A/F43A | -0.3 ± 3.0 | ||
| Thr-47 | iii | T47E | 8.8 ± 1.6 |
| Glu-49 | iii | E49A | 16.3 ± 0.3 |
| T47E/E49A | 42.3 ± 13.1 | ||
| Ala-51 | iii, vi | A51P | 3.5 ± 3.4 |
| T47E/E49A/A51P | -1.1 ± 4.4 | ||
| Lys-56 | i | K56A | 82.1 ± 9.1 |
| Lys-67 | i | K67A/K70A | |
| Lys-70 | i | K67A/K70A | 69.1 ± 6.0 |
| Lys-73 | i | K73A/K74A/K75A | 61.0 ± 6.3 |
| Arg-74 | i | 4K/R>Ad | 14.2 ± 0.5 |
| Lys-75 | i, ii | 5K/R>Ae | -0.4 ± 2.9 |
Human LEDGF/p75 (National Center for Biotechnology Information accession number NP_150091).
i, role in HDGF DNA binding (31); ii, well conserved among PWWP domain orthologs (39, 58) and/or Tudor domain Royal Family members (35) and surface exposed (31, 39); iii, predictive hydrophobic cavity exposure (31, 39); iv, conserved among HRP family members (6, 22, 58) and exposed on the presumptive DNA binding face (Fig. 2B) (31, 39); v, surface exposed near the hydrophobic cavity (Fig. 2C) (31, 39); vi, analogous DNMT3B residue (Ser-270) implicated in ICF syndrome (53).
That is, the percent HIV-Luc activity from two to four independent experiments relative to E6(-/-) cells transfected with wild-type LEDGF/p75 expression vector. Previously described ΔPWWP and ΔPWWPΔATh deletion mutants functioned at 19.2% ± 2.5% and −3.4% ± 4.2%, respectively, whereas the MutL1 NLS/ATh mutant (Fig. 1A) supported 159% ± 29.7% of wild-type activity.
K67A/K70A/K73A/R74A/K75A.
K56A/K67A/K70A/K73A/R74A/K75A.