Tethering of YFP-LacI-G9a-ΔSET or YFP-LacI-G9a-H1166K (a point mutant, which abolishes the catalytic ability of G9a) to the locus failed to recruit CFP-ORCA.
Finally, the FK2 signal at the posterior of the oocyte is not due to the accumulation of ubiquitylated pole plasm components, because it persists in the vas PD mutant, which abolishes pole plasm formation downstream of Osk.
Although we used multiple Bcl-2 deletion mutants covering the entire Bcl-2 sequence, including the ΔBH4/loop mutant which abolishes the binding with SOD1-WT, we failed to identify specific domains in Bcl-2 responsible for binding with mutSOD1.
The phosphorylated USP15 was recruited to DSBs (Fig. 4e and Supplementary Fig. 5e), while the S678A mutant, which abolished its phosphorylation by ATM, could not be recruited to DSBs (Fig. 4e).
To investigate the role of phosphorylation on BCA2 stability, HEK293T cells were co-transfected with a constitutively active variant of AKT or a kinase-dead AKT mutant, along with FLAG-tagged BCA2 or the S132, 133A mutant which abolished AKT phosphorylation.
2. The methyl-lysine-recognizing Agenet domains of FMRP may still needed for its neuronal function, though Alpatov et al. have elucidated that the replication stress defective FMRP mutants, which abolish its methylated histone binding function, do not alter AMPAR internalization.
The similarities between the feelgood phenotype and the phenotypes of the crusher and bulldog mutants, which abolish the function of two key COPII components, suggested that the feelgood locus encodes a protein participating in COPII-mediated transport.
To pursue this model, we substituted R483 in the SH2 pTyr-binding pocket with methionine (mutant R/M), which abolishes pTyr recognition.
Because analysis of the isolated N-terminal module indicated possible disruption of its native structure, we also generated and purified the CBS-1 mutants K421A, which abolishes a canonical PLP-binding site in the C-terminal module, and E62K, which creates a putative PLP-binding site in the N-terminal module.
Hr N mutants harbor the mutation A402G, which abolishes the same uATG as in Hr Hp mutants, indicating that they have an identical defect [ 16].
Similarly, we show that steady-state ELISA experiments conducted with HIF-2αOH (amino acids 523-541) peptides (i.e., hydroxyl group synthetically added to P531) and purified VBC complex suggested that both class 1 and class 2 mutations had negligible or minimal effect on pVHL affinity (Fig. 4a), with the exception of the P531A mutant, which completely abolished binding to VBC as expected.
