Nova autoregulation reveals dual functions in neuronal splicing | The EMBO Journal  Nova autoregulation reveals dual functions in neuronal splicing | The EMBO Journal
RNase T1 digestion (T1) was used to generate size standards. Boundaries are highlighted by arrows and indicated on the ... RNase protection assays and RT-PCR. RNA preparation and RNase protection assays were performed as described previously (Dredge ... To confirm these results, we also performed RNase protection assays (RPA), which revealed a 24.6% decrease in steady‐state Nova ... The pellet was treated with RNase prior to separation by SDS-PAGE, transferred to nitrocellulose and exposed to autoradiography ...
http://emboj.embopress.org/content/24/8/1608
Roles of the influenza virus polymerase and nucleoprotein in forming a functional RNP structure | The EMBO Journal  Roles of the influenza virus polymerase and nucleoprotein in forming a functional RNP structure | The EMBO Journal
Lanes 'C' are incubation controls without RNase. Lanes '1' are RNase T1 digestions of panhandle RNA under native conditions ... RNase T1 digestion. The digestion was carried out in buffer A (see above) with 0, 0.1, 0.5 or 1 U of the enzyme in a 300 μl ... The cleavage by RNase T1 is indicated by arrowheads. The scheme was drawn with the sequence of segment 8 vRNA in the centre. ... RNase T1 digestion of a 3′ end‐labelled, 81 nucleotide panhandle model RNA in the presence of increasing concentrations of DOC ...
http://emboj.embopress.org/content/16/6/1248
Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE‐containing mRNAs | The...  Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE‐containing mRNAs | The...
Total RNA was isolated for T1 RNase protection analyses using both a β‐globin and a CAT antisense probe. (B) Time‐course of β‐ ... Total RNA was isolated for T1 RNase protection analyses using both a β‐globin and a CAT antisense probe. (D) Time‐course of ... C overnight to allow annealing and then digested with T1 RNase (1.2 unit/10 μg RNA, Calbiochem) at 30°C for 1 h. Samples were ... total RNA was isolated and analyzed by RNase T1 protection (Figure 3A). The data, averaged from several experiments and ...
http://emboj.embopress.org/content/17/12/3448
Cooperation of enzymatic and chaperone functions of trigger factor in the catalysis of protein folding | The EMBO Journal  Cooperation of enzymatic and chaperone functions of trigger factor in the catalysis of protein folding | The EMBO Journal
RNase T1 were purified as described (Stoller et al., 1995, 1996; Mücke and Schmid, 1994). (S54G,P55N)‐RNase T1 as well as α‐ ... As in previous work (Stoller et al., 1995), we use a reduced and carboxymethylated variant of ribonuclease T1 (RCM‐T1) as a ... Mayr LM, Odefey C, Schutkowski M and Schmid FX (1996) Kinetic analysis of the unfolding and refolding of ribonuclease T1 by a ... T1 was measured between 0.1 and 10 μM RCM‐T1 at pH 8.0, 15°C. Under these conditions RCM‐T1 does not aggregate, and its ...
http://emboj.embopress.org/content/16/1/54
Sen1 has unique structural features grafted on the architecture of the Upf1‐like helicase family | The EMBO Journal  Sen1 has unique structural features grafted on the architecture of the Upf1‐like helicase family | The EMBO Journal
After incubation for 1 h at 4°C, the reaction mixtures were digested with 1 μg RNase A/T1 mix and 2.5 U RNase T1 (Fermentas) ... mer RNA in the presence of the indicated proteins with RNase A and RNase T1. The left lane was loaded with 10‐mer and 15‐mer ... RNase protection assays with yeast Sen1Hel, Upf1Hel, and Upf1CH‐Hel in the presence of different nucleotides (ADP:BeF3− and ADP ... RNase protection assays. Proteins (10 pmol each) were mixed with 5 pmol 32P body‐labeled RNA to a final 20 μl reaction volume ...
http://emboj.embopress.org/content/early/2017/04/13/embj.201696174
MicroRNA genes are transcribed by RNA polymerase II | The EMBO Journal  MicroRNA genes are transcribed by RNA polymerase II | The EMBO Journal
RNase T1 was used for digestion instead of RNase A/T1 mix. ... Knight SW, Bass BL (2001) A role for the RNase III enzyme DCR‐1 ... Lee Y, Ahn C, Han J, Choi H, Kim J, Yim J, Lee J, Provost P, Radmark O, Kim S, Kim VN (2003) The nuclear RNase III Drosha ... Bernstein E, Caudy AA, Hammond SM, Hannon GJ (2001) Role for a bidentate ribonuclease in the initiation step of RNA ... Ribonuclease protection assay. RPA was carried out as previously described (Lee et al, 2002). Briefly, the whole RNA (100%) ...
http://emboj.embopress.org/content/23/20/4051
RNA‐dependent recruitment of the origin recognition complex | The EMBO Journal  RNA‐dependent recruitment of the origin recognition complex | The EMBO Journal
C) GST or GST-LR1 were incubated with HeLa nuclear extract and then treated with either 0.02 mg/ml RNase A, 2 U/ml RNase T1, ... To determine RNA sensitivity, RNase A (Roche) in a concentration of 0.02 mg/ml or RNase T1 (Roche) in a concentration of 2 U/ml ... We found that ORC recruitment by LR1 can be inhibited by RNase A, RNase T1, and micrococcal nuclease I (MNase I), all of which ... or 100 μg of tRNA under the same conditions for RNase A, RNase TI, DNase I, and MNase I used in the IPs and GST pull‐downs ...
http://emboj.embopress.org/content/27/22/3024
Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination | The EMBO Journal  Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination | The EMBO Journal
Also analysed were the RNA input (In; lane 1), a partial T1 nuclease digest (T1; T1 cuts RNA after guanosines) and a partial ... which targeted RNase H (Figure 3D, lanes 5-11) and formation of distinct cleavage products showed that RNase H cleavage was ... D) RNase H protection analysis with Yhh1p and sCYC1 RNA. Internally 32P‐labelled sCYC1 RNA was incubated with binding buffer ( ... RNase H reactions on total RNA were carried out with oligo ACT1‐RNaseH (CTGGGAACA TGGTGGTACCACC) and RNAs were separated on 8.3 ...
http://emboj.embopress.org/content/21/15/4125
Silencing and transcriptional properties of the imprinted Airn ncRNA are independent of the endogenous promoter | The EMBO...  Silencing and transcriptional properties of the imprinted Airn ncRNA are independent of the endogenous promoter | The EMBO...
E) Sequence showing locations of published Airn (T1) and PGK (T1(PGK)) transcription start sites and two transcription start ... These findings were confirmed by RNase Protection Assay, which showed a complete absence of Airn ncRNA in differentiated APD‐1 ... T1) and downstream CpG island are unchanged. SD: Airn splice donor. A 3.65‐bp PacI-BamHI fragment that has been genetically ... T1) (126236-127195 bp in accession number AJ249895). Constructs 2 and 3 were used to insert the ubiquitously expressed mouse ...
http://emboj.embopress.org/content/27/23/3116
Recognition of polyadenylation sites in yeast pre‐mRNAs by cleavage and polyadenylation factor | The EMBO Journal  Recognition of polyadenylation sites in yeast pre‐mRNAs by cleavage and polyadenylation factor | The EMBO Journal
RNase H mapping: for each assay 0.5-1.0 μg of protein was bound to 50 fmol of labelled RNA in RNase H buffer [100 mM KCl, 50 mM ... U to G changes were visualized by nuclease T1 digestions (T1 nuclease cuts RNA 3′ of Gs; Figure 2A, lanes 6, 11, 16, 21, 26 and ... Also shown are T1 nuclease reactions (T1; lanes 1, 6, 11, 16 and 21), alkaline hydrolysis ladders (OH−; lanes 2, 7, 12, 17 and ... A) RNase H protection profile of purified CPF on sCYC1. The assays were performed with the same substrate and DNA ...
http://emboj.embopress.org/content/20/12/3197
SHLD2/FAM35A co‐operates with REV7 to coordinate DNA double‐strand break repair pathway choice | The EMBO Journal  SHLD2/FAM35A co‐operates with REV7 to coordinate DNA double‐strand break repair pathway choice | The EMBO Journal
Cell pellets were resuspended in 6 ml DNA lysis buffer (10 mM Tris-Cl, 10 mM EDTA, 0.5% SDS, pH 8.0) with 100 μg/ml RNase A, ... Endocr Relat Cancer 23: T1-T17. OpenUrlAbstract/FREE Full Text. *↵. Gaudreau‐Lapierre A, Garneau D, Djerir B, Coulombe F, Morin ... Cells were washed and resuspended in a propidium iodide (PI) solution (20 μg/ml PI, 300 μg/ml RNase, PBS), incubated at RT for ...
http://emboj.embopress.org/content/early/2018/08/28/embj.2018100158
Differential effects of viral silencing suppressors on siRNA and miRNA loading support the existence of two distinct cellular...  Differential effects of viral silencing suppressors on siRNA and miRNA loading support the existence of two distinct cellular...
... triethanolamine and RNase treatments were omitted. Slides were hybridized with ~45 ng of DIG‐labelled probe overnight at 52°C. ... P38 T0 or selected T1 line (#1). The presence of AGO1 in each IP was confirmed by protein blot analysis (upper panel). Total ...
http://emboj.embopress.org/content/31/11/2553?ijkey=0bb9311a40bb385364be28f5bee12c786fda5749&keytype2=tf_ipsecsha
Differential effects of viral silencing suppressors on siRNA and miRNA loading support the existence of two distinct cellular...  Differential effects of viral silencing suppressors on siRNA and miRNA loading support the existence of two distinct cellular...
... triethanolamine and RNase treatments were omitted. Slides were hybridized with ~45 ng of DIG‐labelled probe overnight at 52°C. ... P38 T0 or selected T1 line (#1). The presence of AGO1 in each IP was confirmed by protein blot analysis (upper panel). Total ...
http://emboj.embopress.org/content/31/11/2553?ijkey=aef1816f9748aca8eaa96bc68337164f60038c20&keytype2=tf_ipsecsha
Crystal structure of Cwc2 reveals a novel architecture of a multipartite RNA‐binding protein | The EMBO Journal  Crystal structure of Cwc2 reveals a novel architecture of a multipartite RNA‐binding protein | The EMBO Journal
Pena V, Rozov A, Fabrizio P, Lührmann R, Wahl MC (2008) Structure and function of an RNase H domain at the heart of the ... The complex was hydrolysed by RNases A and T1 (Ambion, Applied Biosystems, Darmstadt, Germany) and protease trypsin (Promega, ... For example, it was recently shown that a C‐terminally located RNase H‐like domain of Prp8 is situated at the heart of the ...
http://emboj.embopress.org/content/31/9/2222?ijkey=5f3c3e0a1ca8a70156816135de5e97721a9e65f4&keytype2=tf_ipsecsha
Aubergine and piRNAs promote germline stem cell self‐renewal by repressing the proto‐oncogene Cbl | The EMBO Journal  Aubergine and piRNAs promote germline stem cell self‐renewal by repressing the proto‐oncogene Cbl | The EMBO Journal
... and either RNase Inhibitor (0.25 U/μl; Promega) or RNase A (0.1 U/μl; Sigma). 50 μl of Dynabeads Protein G (Invitrogen) was ... To address the role of Ago3 in GSC biology, we used ago3 mutant alleles that contain premature stop codons, ago3t1, ago3t2, and ... A-B' Immunostaining of wild‐type (A, A') and ago3t1/t3 mutant (B, B') germaria with anti‐Aub (green) showing that Aub ... ago3t1, ago3t2 and ago3t3 (Li et al, 2009a), nos‐Gal4:VP16 (Rorth, 1998), UASp‐CCR4‐HA (Semotok et al, 2005), UASp‐GFP‐Aub ( ...
http://emboj.embopress.org/content/36/21/3194
Shp2 signaling suppresses senescence in PyMT‐induced mammary gland cancer in mice | The EMBO Journal  Shp2 signaling suppresses senescence in PyMT‐induced mammary gland cancer in mice | The EMBO Journal
For qRT-PCR, total RNA from tumor tissues, freshly isolated single cells, or cultured cells were extracted using TRIzol, RNase‐ ... compare normal tissue with tumors T1‐3). We then undertook a functional analysis of the potential role of Shp2 in the PyMT ...
http://emboj.embopress.org/content/early/2015/03/03/embj.201489004