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LANA R9 is critical in nucleosome binding as mutation of this residue eliminates LANA's chromatin association.
Our data suggest that Ser comprises the critical CDK phosphorylation site controlling PLK1 binding as mutation of Ser, but not Ser or Ser, suppressed interaction of NUAK1 with βTrCP as well as PLK1.
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What is more, the N-terminal conserved residuals are also involved in the DNA binding, such as, mutation of conserved residuals (G182A/F183) located within N-terminal domains of phiC31 integrase had completely lost their ability to bind to the specific target DNA attB.
This regulation occurs through the predicted target miR-4732-3p binding sites, as mutation of the respective target sequences in the 3' UTR of SMAD2 and 4 abrogated the endogenous microRNA-mediated luciferase repression (Fig. 4d).
Mutation of this residue results in a similar increase of peptide-binding affinity as mutation of the fic His [5], [19].
The effect of glycogen and cyclodextrin on the interaction was dependent on their binding to the CBM as mutation of W99, a key residue for cyclodextrin/glycogen binding, greatly blunted the signal reduction.
The exact role of the second SAM remains unknown, but its binding is clearly of physiological significance, as mutation of residues involved in the second SAM binding abolished the coproporphyrinogen III oxidase activity.
In all cell types, CRTR-1 activity is mediated through the CP2 binding elements in the reporter construct, as mutation of these sites abrogates activity (data not shown).
This activation demonstrates dependence on binding of Pbx1 or HoxA9 to its consensus binding sequences in the Hif-1α reporter as mutation of the seed sequences (PbxMut-Hif-1α-pGL2 or HoxMut-Hif-1α-pGL2 reporters, respectively) completely abolished the activation of Hif-1α by Pbx1 or HoxA9 respectively.
This does not appear to be the case during mitosis, as mutation of all CPEB binding sites in the 3′UTR of Emi1 does not prevent translational repression in mitosis (unpublished observation).
Since the interlobar cleft is the site of ATP and carbohydrate binding as well as where phosphoryl transfer occurs, mutation of these residues could change the interlobar movement during catalysis and thus affect the kinase activity of the protein.
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