The thymocyte-specific RNA-binding protein Arpp21 provides TCR repertoire diversity by binding to the 3'-UTR and promoting Rag1 mRNA expression.

The regulation of thymocyte development by RNA-binding proteins (RBPs) is largely unexplored. We identify 642 RBPs in the thymus and focus on Arpp21, which shows selective and dynamic expression in early thymocytes. Arpp21 is downregulated in response to T cell receptor (TCR) and Ca2+ signals. Downregulation requires Stim1/Stim2 and CaMK4 expression and involves Arpp21 protein phosphorylation, polyubiquitination and proteasomal degradation. Arpp21 directly binds RNA through its R3H domain, with a preference for uridine-rich motifs, promoting the expression of target mRNAs. Analysis of the Arpp21-bound transcriptome reveals strong interactions with the Rag1 3'-UTR. Arpp21-deficient thymocytes show reduced Rag1 expression, delayed TCR rearrangement and a less diverse TCR repertoire. This phenotype is recapitulated in Rag1 3'-UTR mutant mice harboring a deletion of the Arpp21 response region. These findings show how thymocyte-specific Arpp21 promotes Rag1 expression to enable TCR repertoire diversity until signals from the TCR terminate Arpp21 and Rag1 activities.

Disrupting Roquin-1 interaction with Regnase-1 induces autoimmunity and enhances antitumor responses.

Roquin and Regnase-1 proteins bind and post-transcriptionally regulate proinflammatory target messenger RNAs to maintain immune homeostasis. Either the sanroque mutation in Roquin-1 or loss of Regnase-1 cause systemic lupus erythematosus-like phenotypes. Analyzing mice with T cells that lack expression of Roquin-1, its paralog Roquin-2 and Regnase-1 proteins, we detect overlapping or unique phenotypes by comparing individual and combined inactivation. These comprised spontaneous activation, metabolic reprogramming and persistence of T cells leading to autoimmunity. Here, we define an interaction surface in Roquin-1 for binding to Regnase-1 that included the sanroque residue. Mutations in Roquin-1 impairing this interaction and cooperative regulation of targets induced T follicular helper cells, germinal center B cells and autoantibody formation. These mutations also improved the functionality of tumor-specific T cells by promoting their accumulation in the tumor and reducing expression of exhaustion markers. Our data reveal the physical interaction of Roquin-1 with Regnase-1 as a hub to control self-reactivity and effector functions in immune cell therapies.

Optically active tetra-tert-butyl-P(5)-deltacyclene epimers: preparation, spectroscopy, dynamic equilibriums, H/D exchange, and transition-metal complex chemistry.

On the basis of isolated diastereomeric triorganylstannyl-P5 -deltacyclenes 7' and 7'', almost pure enantiomers of their destannylation products 8' and 8'' are now available. These stereochemically inert cage chiral species contain a configurationally labile P1H1 group that defines two epimers 8 a and 8 b of each of the enantiomers, which are connected by a rapid equilibrium. Mirror-symmetric circular dichroism (CD) spectra of the enantiomeric cages are compatible with the identification of epimers. A simulation of the CD spectrum of the major epimer 8'a relates the cage chirality of the system to the observed chiroptical effects. Both cage epimers and two of the phosphorus cage atoms are active as ligands with respect to [M(CO)5 ] fragments of Cr, Mo, and W. Four almost isoenergetic regio- and stereoisomers of the resulting mononuclear complexes are formed for these metals, but only one of the isomers per metal crystallized in the case of the racemic series of the complexes. The enantiopure versions of cages and cage complexes, however, did not crystallize at all, a well-known phenomenon for chiral compounds. CD spectra of the optically active complex isomer mixtures are close to identical with the CD spectra of the related free cages and point again to the chiral cages as the dominant source of the CD effects of the complexes. [(Benzene)RuCl2 ] complexes of the cage ligand 8 behave totally differently. Only a single species 12=[(benzene)RuCl2 ⋅8 b] is formed in almost quantitative yield and the minor epimer 8 b plays the role of the ligand exclusively. The reaction works as well for the separated enantiomeric cage versions to yield the highly enriched enantiomers 12' and 12'' separately. An efficient kinetic resolution process was identified as the main reason for this finding. It is based on a high stereo- and regiochemical flexibility of the PC cage ligand that is capable of adjusting to the specific requirements of a suitable transition-metal complex fragment. Such ligand flexibility is regularly observed in metalloenzymes, but is a very rare case in classical and organometallic complex chemistry.

Cage chirality of P-C cage compounds: highly diastereoselective formation of diastereomeric P5-deltacyclenes, separation of diastereomers, and removal of the chiral auxiliary.

An effective cyclic addition reaction of diastereomeric (R*)diphenyltin-3,5-di(tert-butyl)-1,2,4-triphosphole derivatives 6 a-c (R* = (-)-cis-myrtanyl (a), (-)-trans-myrtanyl (b), m-(2-bornyl-2-ene)phenyl (c) with two equivalents of tert-butylphosphaalkyne 1 leads to 1:1 mixtures of diastereomeric stannylated pentaphosphadeltacyclene derivatives 7 a-c with seven stereogenic centers in the cage unit. The (-)-cis-myrtanyl derivative 7 a could be separated into its diastereomers; destannylation of diastereomer 7 a(RR) led to the P-H cage 8 as a pure enantiomer. Circular dichroism (CD) spectroscopy of the pure diastereomer 7 a(RR) and the enantiomer 8 give evidence for identical stereoisomers of the P(5)-deltacyclene cage units and prove a strong dominance of the chiral cages over the chiral auxiliary groups with respect to their chiroptical properties. Absolute X-ray structure investigations of the majority of the compounds presented in the paper reveal the details of the stereochemistry of the asymmetric P-C cage units. In this paper we demonstrated for the first time a general preparative route to stereochemically fully defined asymmetric P-C cage compounds by separation of diastereomers and replacement of the chiral auxiliary group.