Two genes that encode ribosomal-protein S6 kinase homologs are induced by cold or salinity stress in Arabidopsis thaliana

AbstractWe have isolated two closely related cDNA clones (cATPK19 and cATPK6) with homology to protein-serine/threonine kinases from Arabidopsis thaliana using the polymerase chain reaction (PCR). The deduced amino acid sequences of the ATPK19 and ATPK6 contain all 11 conserved regions of the catalytic domain of protein kinases, and have homology to p70 ribosomal S6 kinases (52%). ATPK19 and ATPK6 have putative PEST regions in their N- and C-terminal regions, and these regions also contain putative phosphorylation sites that are recognized by casein kinases or proline-directed protein kinases such as cdc2, MAP kinase, and p54 MAP-2 kinase (SAPK). The transcription levels of the ATPK19 and ATPK6 genes rapidly and markedly increased when plants were subjected to cold or high-salt stresses. These observations suggest that ATPK19 and ATPK6 may function in the adaptation of plant cells to cold or high-salt conditions, providing an understanding of the role of protein phosphorylation in plant responses to environmental stresses

Crystal Structure of a Human K‑Ras G12D Mutant in Complex with GDP and the Cyclic Inhibitory Peptide KRpep-2d

The Ras proteins play roles in cell differentiation, proliferation, and survival. Aberrant signaling through Ras-mediated pathways in tumor cells occurs as a result of several types of mutational damage, which most frequently affects the amino acids G12, G13, and Q61. Recently, KRpep-2d was identified as a K-Ras­(G12D) selective inhibitory peptide against the G12D mutant of K-Ras, which is a key member of the Ras protein family and an attractive cancer therapeutic target. In this study, the crystal structure of the human K-Ras­(G12D) mutant was determined in complex with GDP and KRpep-2d at 1.25 Å resolution. This structure revealed that the peptide binds near Switch II and allosterically blocks protein–protein interactions with the guanine nucleotide exchange factor. This discovery of a unique binding pocket provides valuable information that will facilitate the design of direct Ras inhibitors

Phase I Clinical Trial of Fibronectin CH296-Stimulated T Cell Therapy in Patients with Advanced Cancer

<div><p>Background</p><p>Previous studies have demonstrated that less-differentiated T cells are ideal for adoptive T cell transfer therapy (ACT) and that fibronectin CH296 (FN-CH296) together with anti-CD3 resulted in cultured cells that contain higher amounts of less-differentiated T cells. In this phase I clinical trial, we build on these prior results by assessing the safety and efficacy of FN-CH296 stimulated T cell therapy in patients with advanced cancer.</p><p>Methods</p><p>Patients underwent fibronectin CH296-stimulated T cell therapy up to six times every two weeks and the safety and antitumor activity of the ACT were assessed. In order to determine immune function, whole blood cytokine levels and the number of peripheral regulatory T cells were analyzed prior to ACT and during the follow up.</p><p>Results</p><p>Transferred cells contained numerous less-differentiated T cells greatly represented by CD27+CD45RA+ or CD28+CD45RA+ cell, which accounted for approximately 65% and 70% of the total, respectively. No ACT related severe or unexpected toxicities were observed. The response rate among patients was 22.2% and the disease control rate was 66.7%.</p><p>Conclusions</p><p>The results obtained in this phase I trial, indicate that FN-CH296 stimulated T cell therapy was very well tolerated with a level of efficacy that is quite promising. We also surmise that expanding T cell using CH296 is a method that can be applied to other T- cell-based therapies.</p><p>Trial Registration</p><p>UMIN <a href="https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr.cgi?function=brows&action=brows&type=summary&recptno=R000002101&language=J" target="_blank">UMIN000001835</a></p></div

Correlation between the number of less-differentiated T-cell surface markers of transferred cells (after culture) and those of PBMCs (before culture).

<p>The comparison was done in terms of cell-surface markers (i.e. CD27+CD45RA+, CD28+CD45RA+, CCR7+CD45RA+).</p

Characteristics of infused cells.

<p>Patient no. 1 to 6 needed a one-time culture and patients 7 to 9 underewent three lymphocyte cultures.</p><p>Values are expressed as mean±SD.</p