IDENTIFYING THE PATH OF THE RNA PRIMER AROUND A SIMPLE RNA POLYMERASE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 牛痘病毒多聚腺苷酸聚合酶（VP 55）是已知的唯一相对于其RNA引物移位的RNA聚合酶，这引起了一个悖论：聚合酶或引物哪个更灵活？该蛋白质有两种已知的作用模式，即进行性合成的30 nt的尾巴，并作为异源二聚体与其持续合成因子（VP 39），半进行性合成的200？300新台币的尾巴。过去的合作研究已经产生了VP 39和VP 55-VP 39异源二聚体的晶体结构。最重要的，但缺乏的信息是异源二聚体引物复合物的结构理解。通过使用同事的光束时间，格申实验室的全职晶体学家在寻找聚合酶表面短RNA片段的结合位点方面取得了显着进展。虽然这是一个开创性的，原始的工作聚（A）聚合酶，3小时的光束时间每2个月提供了不充分的进展，在这个NIH资助的工作，以确保赠款更新。射束时间是主要的瓶颈，因此，目前的要求，个人，定期射束时间的结晶工作。

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. Vaccinia virus poly(A) polymerase (VP55) is the only RNA polymerase known to translocate with respect to its RNA primer, raising a paradox: Which is more flexible: The polymerase or the primer? The protein has two known modes of action, namely the processive synthesis of 30 nt tails and, as a heterodimer with its processivity factor (VP39), the semi-processive synthesis of 200 ? 300 nt tails. Past, collaborative studies, have yielded crystal structures for VP39, and for the VP55-VP39 heterodimer. The most important, yet absent information is a structural understanding of the heterodimer-primer complex. By using the beam time of a colleague, a full-time crystallographer in the Gershon lab has progressed markedly in finding binding sites for short RNA segments on the polymerase surface. Although this is ground-breaking, original work for a poly(A) polymerase, 3 hours of beam time every 2 months provides inadequate progress in this NIH-funded work to ensure grant renewal. Beam time is the major bottleneck, hence the current request for individual, regular beam time for this crystallographic work.