Trypanosome SF3a60 localizes in the nucleus.

<p>(A) Cells expressing <i>SF3a60</i>-TAP and the TAP tag with (+) and without (−) tetracycline induction after Western blot analyses. The SF3a60-TAP is conditionally expressed while tubulin (TUB) expression is used as control and (B). Immunofluorescence of procyclic form trypanosomes TAP-tagged <i>TbSF3a60</i> counterstained for <i>Tb</i>SF3a60 (red) and for DNA (DAPI).</p

SF3a60-associated protein preys from a genome-wide Yeast-2-hybrid screen.

1<p>Confidence levels according to PBS scores <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091956#pone.0091956-Formstecher1" target="_blank">[32]</a>.</p>2<p>Domains (superfamily, Pfam) predicted in TritrypDB (<a href="http://tritrypdb.org/tritrypdb/" target="_blank">http://tritrypdb.org/tritrypdb/</a>); a blank space indicates those without known domains.</p>3<p>Functional designation. Functions predicted on the basis of sequence match alone are shown in parentheses.</p

Sequence conservation for putative T. brucei PRP40.

<p><b>A</b>. The WW domains of human FBP11 (O75400.2), yeast PRP40 (NP_012913.3) and the putative Trypanosome FBP11 homologue contain the tryptophans, prolines and a central set of three hydrophobic residues characteristic of WW domains. Evidently, the putative <i>T. brucei</i> FBP11 lacks a second WW domain.</p

S3a60 is essential for trypanosome viability.

<p>(A) Cumulative growth curves of TbSF3a60 depleted trypanosomes. The cells grown with (triangles, solid line) or without (squares, dashed line) tetracycline. Every 24 h, samples were taken for the determination of cell density (cells/ml), and grown cultures were diluted down to 5×10⁵ cells/ml with fresh medium. (B). Effect on <i>SF3a60</i> mRNA. Cells were grown either without <i>Tet</i> (–), or in the presence of 100 ng/ml <i>Tet</i> (+) for 24, 48 and 72 hours. Each lane on the Northern blot contains 10 μg RNA from bloodstream form wild type (WT) or an RNAi cell line. Asterisk (*) denotes dsRNA. Trypanosome SRP and Tubulin (TUB) RNAs are used as loading controls. Arrow indicates SF3a60 mRNA.</p

Preparative tandem affinity purification identifies SF3a60 associated proteins.

<p>Proteins were separated on 12% SDS-PAGE and stained with SYPRO Ruby. Marker protein sizes in kilo Daltons are indicated on the left and corresponding protein bands are on the right. Details are in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091956#pone-0091956-t001" target="_blank">Table 1</a>.</p

SF3a60 RNAi has no effect on splicing.

<p>The Y structure <i>trans</i> splicing intermediate is not decreased after two days of RNAi targeting SF3a60. The full-length spliced leader RNA and the Y structure were detected by primer extension (diagram in panel A) followed by denaturing polyacrylamide gel electrophoresis (B) with the U3 snRNA as a loading control. The RNA was prepared from the RNAi cell line grown without tetracycline (panel B, lanes 1 and 4), or with tetracycline for one or two days (lanes 2 and 3). M: markers.</p

Proteins associated with SF3a60 by tandem affinity purification.

<p>Proteins associated with SF3a60 by tandem affinity purification.</p

Structure of the Fab17.2 – R13 complex.

<p>A. Superposition of the apo Fab 17.2 and R13 complex structures (grey and green respectively). VH and VL contact residues are indicated. B. Main water molecules present on the antigen binding site of Fab 17.2 apo. C. Superposition of water molecules present in the apo Fab 17.2 that are replaced by the peptide in the Fab 17.2 R13 complex. D. Superposition of R13 peptides from molecules 1 (green) and 2 (magenta). E. Structure of the Fab 17.2-R13 complex (molecule 1). All hydrogen bonds between mAb 17.2 and peptide R13 are illustrated as dotted yellow lines. The π-stacking interaction between VL Tyr101 and the epitope Phe9 is also indicated. Heavy chain CDRs are coloured in red and light chain CDRs in blue the peptide is coloured in light green.</p

Polar contacts in the model between 17.2 and β1-AR.

<p>MC: Main Chain; SC: Side Chain.</p

Crystallographic data and refinement statistics.

<p>Crystallographic data and refinement statistics.</p