One unit of M-MLV reverse transcriptase (Promega, M170B) was used as positive control. The MS2 reverse-transcribed DNA (cDNA) in samples was quantitated by qPCR with MS2 specific primers (Supplementary information, Table S1). The relative RTase activities between samples were compared based on...
RNA-associated proteins were purified following a modified version of the published TRAPP protocol, where silica columns were used in place of silica beads37. Proteins were digested on the column with 0.25 µg of Trypsin/Lys-C protease mix (Promega; cat. V5071), and peptides eluted for...
In spite of these similarities, RNase A members have not been able to completely replace RNase T2 functions in vertebrates, since at least one gene belonging to the latter family has been found in each animal genome completely sequenced. To gain insights on the evolution and coexistence of ...
One possible explanation for the different RNA specificities of onconase observed in cells and in cell lysates from that of purified RNAs could be that cellular rRNA is bound to proteins and thus protected from onconase in cells and cell lysates, whereas the tRNA is more exposed. Purified rRNA,...
If this were true, however, then one would conclude that a putative late activation of another antiviral pathway also has little relevance in limiting the replication of these viruses. We do not know why HAV/HM175 clone 1 is unable to activate the 2-5OAS/RNase L pathway even after several...
The rice genome encodes three RNase Z proteins, including one long form and two short forms (RNase ZS1 and RNase ZS2; Supplementary Fig. 7). RNase ZS2 localizes in the chloroplast and is essential to chloroplast development31. In humans, ELAC2 (RNase ZL) localizes in the nucleus and ...
Ire1 is activated in response to accumulation of misfolded proteins within the endoplasmic reticulum as part of the unfolded protein response (UPR). It is a unique enzyme, possessing both kinase and RNase activity that is required for specific splicing o
One cDNA clone, named rcr gene, contained an 883-nucleotide insert including a 31-base poly(A) tail. It had a single open reading frame with the first ATG initiation codon located at nucleotide 244 from the 5′ end of the clone followed by a 399-nucleotide open reading frame and ending...
Total RNA from cultured cells was extracted with Maxwell® RSC simplyRNA Tissue Kit with the Maxwell® RSC Instrument (Promega), according to manufacturer instructions. For dilncRNAs detection, chromatin-bound RNA was extracted as follows. Cells were fractionated according to a published protocol65...
The inflammasome is a multiprotein complex assembled in response to Pathogen Associated Molecular Patterns (PAMPs) and Danger Associated Molecular Patterns (DAMPs). Inflammasome activation occurs through a two-step mechanism, with the first signal facili