Disease relevance of POLR3E

• When total HIV (PLB) RNA was used as the source of primer/template, and similarly extended with RT in the presence of radioactive dCTP, the major priming tRNA resolved by RPC-5 had a chromatographic mobility identical to peak 3 [1].
• It was proved by RPC-5 chromatography of tRNA Val that Val-RS I and Val-RS II recognized all five isoacceptor tRNA Val species both from wheat germ and leaves, as well as tRNA Val from E. coli [2].

High impact information on POLR3E

• Here we show that alterations in the LRS domain do not result in Sin(-) phenotypes, nor does disruption of the SIN domain lead to loss of ribosomal DNA heterochromatic gene silencing (Lrs(-) phenotype) [3].
• The LRS and SIN Domains: Two Structurally Equivalent but Functionally Distinct Nucleosomal Surfaces Required for Transcriptional Silencing [3].
• Genetic experiments have identified two structurally similar nucleosomal domains, SIN and LRS, required for transcriptional repression at genes regulated by the SWI/SNF chromatin remodeling complex or for heterochromatic gene silencing, respectively [3].
• In contrast to these dissimilarities, we find that the SIN and LRS domains are both required for recruitment of Sir2p and Sir4p to telomeric and silent mating type loci, suggesting that both surfaces can contribute to heterochromatin formation [3].
• Our results provide a characterization of human RNA polymerase III and show that the RPC5 subunit is essential for transcription [4].

Biological context of POLR3E

• SIN, a novel Drosophila protein that associates with the RNA binding protein sex-lethal [5].
• Separating the gag-pol genome into two distinct expression cassettes (gag-pro and vpr-RT-IN) may reduce the potential for RCL formation, while concurrently employing cSIN vectors supports retention of the SIN phenotype in target cells and alleviates technical constraints associated with generating producer cell lines [6].
• We concluded that the CRT strategy used in the context of SIN replicon particles facilitated the generation of a highly effective vaccine for cancer prophylaxis and immunotherapy [7].
• These results suggest that the nitric oxide-containing SIN 1 inhibits platelet aggregation not only in vitro but also in vivo and that this property can be a beneficial effect in antianginal therapy [8].

Anatomical context of POLR3E

• RPC-5 chromatography separated four major isoacceptors from each source, with those from HeLa cells eluting generally later than those from placenta [9].
• Polyribosome-bound and -unbound isoaccepting species of tRNALys and tRNAIle, isolated from lupin cotyledons, were compared by RPC-5 chromatography and it was found that polyribosomes preferentially bind some of the isoaccepting species [10].
• A tRNAAsn could be charged in isolated CHO TK- cell mitochondria and the asparaginyl-tRNA was found to elute before its cytosolic counterpart on an RPC-5 column and to have a higher mobility on polyacrylamide slab gels run under denaturing conditions [11].

Associations of POLR3E with chemical compounds

• The key steps in this procedure were the joining of dCk oligomers, protected at the 3'-OH with an acetyl group, to rCi oligomers by T4 DNA ligase and the purification of the products by RPC-5 column chromatography [12].
• Individual isoacceptors were separated by chromatography on RPC-5 and then acylated with the 3-(4-hydroxyphenyl)propionyl ester of N-hydroxysuccinimide [13].
• Subsequent chromatography of the partially purified tRNA using high-speed, high-pressure liquid chromatography on RPC-5 and Aminex A-28 coupled with chromatography on BD-cellulose at acidic pH and on DEAE-Sephadex A-50 significantly shortened isolation time for milligram quantities of several pure tRNA species [14].
• Subtraction of the resulting background signal provides NO2 measurements with a limit of detection of 150 ppt/10 s (SIN = 3) [15].
• They can be detected through the use of either RPC-5 column chromatography of tRNA aminoacylated with radioactive lysine or northern blot analysis using a tRNALys3-specific DNA hybridization probe [1].

Analytical, diagnostic and therapeutic context of POLR3E

• High pressure liquid chromatography on the RPC-5 reversed-phase ion exchange system has been shown to have several potential applications as an initial high capacity step in the isolation of specific DNA restriction fragments [16].
• Reactive column HPLC: rapid analysis on RPC-5 of oligoadenylates that differ at the 3'-terminus [17].
• Both RPC-5 chromatography and northern blot analysis show the cytoplasmic form of tRNALys3 to be the major abundance form of tRNALys3 in the virus [1].

References