Biomolecular condensates formed by phase separation are widespread and play critical roles in many physiological and pathological processes. cGAS-STING signaling functions to detect aberrant DNA signals to initiate anti-infection defense and antitumor immunity. At the same time, cGAS-STING signaling must be carefully regulated to maintain immune homeostasis. Interestingly, exciting recent studies have reported that biomolecular phase separation exists and plays important roles in different steps of cGAS-STING signaling, including cGAS condensates, STING condensates, and IRF3 condensates. In addition, several intracellular and extracellular factors have been proposed to modulate the condensates in cGAS-STING signaling. These studies reveal novel activation and regulation mechanisms of cGAS-STING signaling and provide new opportunities for drug discovery. Here, we summarize recent advances in the phase separation of cGAS-STING signaling and the development of potential drugs targeting these innate immune condensates.
Humans ; Nucleotidyltransferases/chemistry* ; Signal Transduction/physiology* ; Membrane Proteins/chemistry* ; Phase Separation

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

AIMTo provide molecular evidence for quality evaluation and GAP production of Pogostemon cablin (Blanco) Benth. cultivated in different regions in Guangdong and Hainan provinces, China, by comparing two sequences (1.2 kb of plastid matK gene and 1.8 kb of nuclear 18S rRNA gene) and two chemotypes (Pogostone-type and Patchouliol-type in essential oil composition).

METHODSPCR direct sequencing was applied to detemine the matK and 18S rRNA sequences for six samples of Pogostemon cablin from different localities.

RESULTSThe matK sequences of six samples of Pogostemon cablin from different regions of cultivation are 1,245 bp in length, which coding 415 amino acids of protein (maturase), and 18S rRNA sequences are 1,803-1,805 bp in size. Based on multiple sequence alignment, there are 47 variable sites in the matK sequence of these six samples, 17 in the 18S rRNA sequence. The cluster tree reconstructed by UPGMA method shows that the sequence divergence both in matK and 18S rRNA genes among six samples of Pogostemon cablin was well correlative with their regions of cultivation and intraspecific chemotypes of essential oil composition.

CONCLUSIONCombining with chemical and biogeographical data, DNA sequencing can become a powerful tool in the key technique-species identification of quality evaluation and GAP production of Pogostemon cablin.

Base Sequence ; DNA, Plant ; analysis ; Endoribonucleases ; genetics ; Genes, Plant ; Lamiaceae ; chemistry ; classification ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Oils, Volatile ; analysis ; Polymorphism, Genetic ; Quality Control ; Sequence Analysis, DNA ; Species Specificity

The systematic position of Fritillaria hupehensis has been in dispute. Phylogentic analyses were conducted on sequences of ITS, rpl16, matK sequences for species of F. hupehensis and allies. Lilium davidii was designed as outgroup. The analyses were performed using MP and ML methods. Conclusions could be achieved as follow. The topologies of MP and ML are consistent. The samples of F. hepehensis from different places form a supported clade with a strong bootstrap. And then form a strongly supported clade with F. anhuiensis, F. monantha. The results suggests that although F. hupehensis has a closet relation with the two ones, it exists some difference.
DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Endoribonucleases ; genetics ; Fritillaria ; classification ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Phylogeny ; Plant Leaves ; genetics ; Ribosomal Proteins ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo study the phylogeny relationship and molecular identification of 10 species from Huperzia (Huperziaceae) based on matK gene sequences data.

METHODTotal DNA of nine species from Huperzia was extracted; matK gene sequence was amplified by PCR. PCR product was directly sequenced after purification.

RESULTThe chloroplast matK gene nucleotide sequences from 9 species of Huperzia species were sequenced. The matK gene nucleotide sequences length was 1 589 bp. Analysis with Huperzia lucidula matK gene nucleotide sequences (download from GenBank) and taking Lycopodiella cernua as outgroup, Maximum Parsimony, Neighbor-Joining analyses and genetic distances were conducted using MEGA 3.1 software. 35 variable sites and 35 parsimony informative sites have been found. Pairwise genetic distances among 10 species of Huperzia was 1.59% - 0.25%.

CONCLUSIONThe results were consistent with the taxonomy in morphological of Huperzia. But H. longipetiolata and H. serrata were resolved into in different clade. There are 19 different sites of matK gene sequences between H. longipetiolata and H. serrata, the genetic distances is 0.121%. It is suggested H. longipetiolata should be as an independent species.

DNA, Chloroplast ; genetics ; DNA, Plant ; chemistry ; genetics ; Endoribonucleases ; genetics ; Huperzia ; classification ; enzymology ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; enzymology ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Species Specificity

In addition to DNA repair pathways, cells utilize translesion DNA synthesis (TLS) to bypass DNA lesions during replication. During TLS, Y-family DNA polymerase (Polη, Polκ, Polı and Rev1) inserts specific nucleotide opposite preferred DNA lesions, and then Polζ consisting of two subunits, Rev3 and Rev7, carries out primer extension. Here, we report the complex structures of Rev3-Rev7-Rev1(CTD) and Rev3-Rev7-Rev1(CTD)-Polκ(RIR). These two structures demonstrate that Rev1(CTD) contains separate binding sites for Polκ and Rev7. Our BIAcore experiments provide additional support for the notion that the interaction between Rev3 and Rev7 increases the affinity of Rev7 and Rev1. We also verified through FRET experiment that Rev1, Rev3, Rev7 and Polκ form a stable quaternary complex in vivo, thereby suggesting an efficient switching mechanism where the "inserter" polymerase can be immediately replaced by an "extender" polymerase within the same quaternary complex.
Binding Sites ; Crystallography, X-Ray ; DNA Repair ; DNA-Binding Proteins ; chemistry ; genetics ; metabolism ; DNA-Directed DNA Polymerase ; chemistry ; genetics ; metabolism ; Fluorescence Resonance Energy Transfer ; Humans ; Mad2 Proteins ; Nuclear Proteins ; chemistry ; genetics ; metabolism ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Protein Binding ; Protein Structure, Quaternary ; Protein Structure, Tertiary ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics