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Interestingly, the elongation promoting activity of Npl3 is weaker in an RNA binding mutant and is inhibited by Rna15, which suggests that binding to the RNA transcript by Npl3 is important for stimulating RNAP II.
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Specifically, we demonstrate significantly increased interaction of both Rab7 disease mutants with ORP1L, a well-characterized effector of active Rab7 that facilitates retrograde trafficking of late endosomes (36), and Vps13C, which also shows increased binding to the Q67L mutant and is likely a mediator of active Rab7 function.
We found that although the WT and the ΔWR TWIST1 mutant were associated with the IL8 promoter, the S144R/K145E DNA binding mutant is not presented at the same region, confirming that these point mutations indeed disrupt the DNA binding ability of the protein.
A typographic error for the YXXΦ binding mutant is present in previous publications (Honing et al., 2005; Edeling et al., 2006; Motley et al., 2006).
An RNA binding mutant was generated by changing two arginines that are essential for RNA binding region to alanines (Tat R52/53A).
The advantage of using this particular RNA-binding mutant is that Flag-TDP-43 F4L is unable to downregulate the endogenous TDP-43 levels by the autoregulation mechanism (6).
Attenuation from the vit A2 consensus ERE is not necessarily dependent on DNA binding as the TR alpha DNA binding mutant was still able to inhibit E-dependent transactivation.
CP-31398, a styrylquinazoline, restores a wild-type DNA-binding conformation of mutant p53, and is able to suppress tumour growth in vitro and in vivo.
Both docking studies led to predicted interface structures consistent with known mutant binding properties and were used to select residues for site-directed mutagenesis.
Both actin binding mutants were expressed and purified in the context of the his6-TEV-KCK-VASP (1 380aa) Cys-light (C7S, C64S, C334A) vector.
Mavs −/− MEF cells reconstituted with WT or TRAF-binding mutant MAVS were infected with Sendai virus for the indicated time.
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