Journal: G3: Genes | Genomes | Genetics

Article Title: New mutations in the core Schizosaccharomyces pombe spindle pole body scaffold Ppc89 reveal separable functions in regulating cell division

doi: 10.1093/g3journal/jkae249

Figure Lengend Snippet: Live-cell imaging of cells expressing Sid4-GFP and Sad1-mCherry in ppc89 + , ppc89-3 , or ppc89-4 cells. Cells were grown at 25°C and were shifted to 36°C for 3 h and images were acquired at both time points. Scale bar, 5 µm. a) Quantification of SPB fluorescence intensity from cells as in (b) and S2B. Sid4-GFP SPB fluorescence intensity was divided by Sad1-mCherry SPB fluorescence intensity. N ≥ 30 cells for each condition and strain from 3 independent experiments. From 3 independent experiments. Error bars represent mean ± SEM. P -values were obtained with a 1-way ANOVA with Tukey's post hoc test. **** P ≤ 0.0001. b) Representative images from the indicated strains. (c, e) Yeast 2-hybrid analysis was performed with S. cerevisiae strain PJ69-4A which was cotransformed with plasmids expressing the indicated Ppc89 and Sid4 fragments. Black boxes indicate regions of predicted α-helices. leu + trp + transformants were tested for growth on −his and -ade plates (SDMU) or -his plates (SDMUA). “+++” indicates strong growth, “++” indicates little growth, and “−” indicates no growth. d) AlphaFold3 predicted structure of the Ppc89-Sid4 complex. Two copies of Ppc89s residues 647-687 were modeled with 2 copies of Sid4 residues 546-660. Ppc89 is in magenta and Sid4 is in gray. f) In vitro binding assay with bead-bound GST or GST-Ppc89(545-687) and soluble MBP or MBP-Sid4(546-660). Samples were washed, resolved by SDS-PAGE, and stained with Coomassie blue.

Article Snippet: Protein structure predictions were generated with the AlphaFold3 (AF) server ( Abramson et al. 2024 ) and visualized using the PyMOL molecular graphics system version 3.0 Schrodinger, LLC.

Techniques: Live Cell Imaging, Expressing, Fluorescence, In Vitro, Binding Assay, SDS Page, Staining

Journal: Infection and Immunity

Article Title: Chlamydia plasmid-encoded protein Pgp2 is a replication initiator with a unique β-hairpin necessary for iteron-binding and plasmid replication

doi: 10.1128/iai.00602-24

Figure Lengend Snippet: Three-dimensional structure of C. trachomatis L2 Pgp2. AlphaFold3 predicted the structure.

Article Snippet: The amino acid sequences of Pgp2 from Chlamydia species, retrieved from the National Center for Biotechnology Information database, were used as inputs for structural prediction using the online AlphaFold3 servers ( ).

Techniques:

Journal: Infection and Immunity

Article Title: Chlamydia plasmid-encoded protein Pgp2 is a replication initiator with a unique β-hairpin necessary for iteron-binding and plasmid replication

doi: 10.1128/iai.00602-24

Figure Lengend Snippet: Conservation of the β-hairpin motif in Pgp2s of all Chlamydia species. ( A ) The amino acids constituting the β-hairpin in C. trachomatis Pgp2 are conserved in all 12 other species. Alignment of Pgp2s of all 13 Chlamydia species was performed using Clustal Omega, although only the 11 amino acid sequences that form the β-hairpin are presented. ( B ) Pgp2s of all 12 other Chlamydia species share the β-hairpin structure (green) with C. trachomatis . All structures were predicted using AlphaFold3.

Article Snippet: The amino acid sequences of Pgp2 from Chlamydia species, retrieved from the National Center for Biotechnology Information database, were used as inputs for structural prediction using the online AlphaFold3 servers ( ).

Techniques: