STRUCT STUD DNA-LESION BYPASS AND EUKARYOTIC TRANSLATIONAL REGULATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 DNA聚合酶kappa(Polk)已被证明可以绕过某些DNA损伤，并从抑制正常DNA合成的未修复错配中延伸出来。我们已经解决了蛋白质的脱氧核糖核酸和与DNA的络合物的结构分别为2.40埃和3.05埃。我们最近表征了一种新的Polk-DNA络合物的晶体形式，如上面的样本1所述，我们乐观地认为，晶体中的新填充安排将使我们能够收集与该共络合物的亚3埃数据。这些数据将使我们能够进行进一步的实验，以了解Polk活性位点如何适应各种DNA扭曲。翻译抑制是真核生物中一种高度保守的基因调控机制，尤其是在胚胎发育过程中。我们已经解决了参与这种抑制的两种蛋白质的结构--人类Pumilio和果蝇Smaug。CNOT6作为CCR4-非死烯基化复合体的一个亚单位参与翻译抑制。为了了解CNOT6在mRNA降解中的作用，我们获得了上面样品2中描述的晶体。这些晶体正在进一步优化，以获得更高分辨率的数据。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. DNA polymerase kappa (Polk) has been shown to bypass certain DNA lesions and to extend from the unrepaired mismatches that inhibit normal DNA synthesis. We have solved the structure of the apo-form of the protein, as well as the co-complex with DNA to 2.40 and 3.05 angstroms respectively. We have recently characterized a new crystal form of the Polk-DNA complex, as outlined in specimen 1 above, and we are optimistic that the new packing arrangement within the crystal will allow us to collect sub-three angstrom data with the co-complex. These data will allow us to pursue further experiments to understand how the Polk active site accommodates a variety of DNA distortions. Translational repression is a highly conserved mechanism of gene regulation in eukaryotes, especially during embryogenesis. We have solved the structure of two proteins involved in such repression -- human pumilio and drosophila smaug. CNOT6 is involved in translational repression as a subunit of the CCR4-NOT deadenylation complex. To understand the role of CNOT6 in mRNA degradation, we have obtained the crystals described in specimen 2 above. These crystals are being further optimized to obtain higher resolution data.