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Table 3 Dominant-negative mutants of TnsD.

From: Characterization of the TnsD-attTn7 complex that promotes site-specific insertion of Tn7

From dominant-negative TnsD screena Relative transposition in presence of Relative attTn7 binding in vivod Relative attTn7 binding in vitro
TnsD proteins TnsABCb TnsABCc   
No TnsD 1.00 0 < 0.00001 -
TnsD WT 1.00 1.13 1.0 1.0
TnsD A18T 0.05 0.01 - -
TnsD P82L 0.05 0.01 - -
TnsD M96I 0.10 0.30 0.005 0.3
TnsD G109E 0.07 < 0.001 0.0006 0.05
TnsD P147L 0.05 < 0.001 0.0016 0.07
TnsD R276C 0.04 < 0.001 - -
TnsD H287Y 0.06 < 0.001 - -
TnsD P417L 0.08 0.04 0.0014 0.07
TnsD C447Y 0.10 0.40 0.012 0.7
TnsD A486H 0.06 0.04 - 0.1
TnsD A497I 0.06 0.04 0.0005 0.05
Truncations     
TnsD Q227Z 0.05 < 0.001 - -
TnsD Q305Z 0.03 < 0.001 - -
TnsD Q306Z 0.03 < 0.001 - -
TnsD Q340Z 0.04 < 0.001 0.0004 < 0.05
TnsD W371Z 0.08 < 0.001 - < 0.05
  1. aThese substitutions and truncations were isolated as dominant negatives and then analysed for transposition in vivo and DNA binding in vivo and in vitro.
  2. bTn7 transposition frequency in vivo was measured using the lambda hop assay and carried out in presence of TnsABCD + TnsD (dominant-negative mutants).
  3. cTn7 transposition frequency in vivo was measured using the lambda hop assay and was carried out in presence of TnsABC + TnsD (dominant-negative mutants).
  4. dThe binding of mutant TnsDs to attTn7 in vivo was measured using the challenge phage assay.