Question

helppp explain the graphs what hypothesis were being made ? what are the controls and conclusion of what is going on

Wee1 Cdc2.3w Cdc2.1w Mik1 G146 148A Cdc25 Cdc2 2.3w 167 T167 CAK T-Loop E FEDCB A - ◆WT kDa O 60- cdc2.1ww 50F wee 30P cdc2.1w 0HM 75HM PT 37.5:1 29:1 A/BA

what is each figure trying to tell us

Answer 1

Based on the given figures let me try to explain what is going on.

In Figure A, wee1 phoshphorylates the Tyrosine (Y) and Theronine (T) residues of Cdc2. The (-) sybmol near Cdc2.1w represents insensitvness to inhibition by Wee and (|--Cdc25) means that, Cdc2.3w does not require activation by Cdc25 phosphotase.Mim1 seems to be involved in modifying the role oh Y15 in Cdc2 here. T167 is also a phosphorylation site and here Wee1 modifies T14 and Y15 and T167 is modified by CAK (Kinases) respectively.

Figure B and C represents two modeled structures of Cdc2.3W and Cdc2.1W. Here, the sites of T14, Y15 and T167 are shown as well as the N-terrminal region and T-loop region is labelled in Cdc2.3W. G(Glycine) 146 seems to be mutated. An ion (yellow sphere) is also located in both the models.

Figure D represents results of SDS page where kDa row shows the marker cloumn and the lower sized bands appear below. PT represents phosphotags and in first gel, only a single band is observed whereas when phophotags are included three different bands H.M and U and observed at different sizes in the both the samples (Cdc2 and Cdc2.1W). The bands are sized at 55kDa, 50kDa and 35 kDa approximately which means that, one of them probably H is fully phosphorylated, M is partially and U in Un phosphorylated protein. So three different forms exist for Cdc2.1W

Figure E represents results of Iso Electric focussing (IEF) on a linear pH gradient ranging from 3-10. Here WT stands generally for wild type protein and $\Delta$Wee1 stands for mutated protein. Due to the mutation in Glycine, there is a shift in the mutant sample towards the positive anode in Cdc2.1W when compared to wild type. Its also noted that this is reversed in $\Delta$Wee1 and Cdc2.1W + $\Delta$Wee1. Here there is a pull towards negative cathode again.

From this, we can infer that three different forms exist for Cdc2.1W and probably the mutant in Gly146 is a negatively charged amino acid (like D- Aspartate) which is then neutralized by Wee1 modification which is again a hypothetical quesion here..Does Cdc2.1w gets modified by Wee1?