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Biology subjects

Li, Y.-z.

Publications and source records attributed to Li, Y.-z..

6 recordsLinked to original sources

Spatial specialization of antioxidant defenses dictates predation success in Myxococcus xanthus

Redox-based chemical warfare is a primary driver of microbial community assembly. Here, we show that the predatory bacterium Myxococcus xanthus employs a spatial division of labor between two inducible monofunctional catalases, mxKatB and mxKatE, to overcome prey-derived hydrogen peroxide (H2O2). Quantitative transcript analysis revealed distinct regulatory specificities: mxkatB was the dominant transcriptional responder to exogenous H2O2, whereas mxkatE was preferentially induced by UV irradiation. Biochemical analyses demonstrated strict compartmentalization of enzymatic activity. mxKatE functioned intracellularly, consistent with a role in mitigating endogenous genotoxic stress. In contrast, mxKatB, which harbors an N-terminal Sec-dependent signal peptide, was exclusively localized to the extracellular milieu. Targeted gene deletions corroborated these non-redundant physiological roles. {Delta}katE mutant exhibited severe growth defects and heightened sensitivity to UV and H2O2 yet retained full predation proficiency. Conversely, {Delta}katB mutant displayed unaltered vegetative fitness but were severely impaired in prey lysis due to oxidative inactivation of secreted bacteriolytic enzymes. Failure of cross-complementation confirmed that spatial localization, rather than catalytic capacity, dictates enzyme function. Our findings establish that M. xanthus deploys an extracellular catalase shield to protect its exoenzyme arsenal from prey-derived oxidants. This spatial specialization of antioxidant defenses represents a sophisticated strategy that directly determines the outcome of bacterial predation and shapes interspecies interactions within microbial communities. IMPORTANCEPredatory bacteria such as M. xanthus must withstand chemical defenses deployed by their prey. We show that M. xanthus uses a spatially specialized antioxidant system: an extracellular catalase (mxKatB) secreted to shield its lytic enzymes from prey-derived hydrogen peroxide, and an intracellular catalase (mxKatE) that handles endogenous oxidative stress. This division of labor reveals that bacterial antioxidant defenses can be compartmentalized to protect extracellular weaponry rather than the cell itself, adding a new dimension to how spatial organization of stress responses influences the outcome of microbial competition.

ecology↗

DnaK duplicates and regionally evolves for the increase of proteomic complexity in bacteria

Hsp70 is important for organismic cells to maintain proteostasis and the chaperone protein is duplicated in all eukaryotes and many prokaryotes. Although the functioning mechanism of Hsp70 has been clearly illuminated, the chaperone duplication and functional evolution has been less investigated. DnaK is a highly conserved bacterial Hsp70 family. Here we showed that the dnaK gene is present in 98.9% bacteria and 6.4% bacteria possess duplicated dnaKs; the occurrence and duplication is positively corelated to the increase of proteomic complexity. We identified the interactomes of the two DnaK paralogs in Myxococcus xanthus DK1622, which are mostly nonoverlapped, but both preferring the &{beta} domain proteins. Consistent with the proteomes, the MxDnaK substrates are both significantly size-larger and pI-higher than that of the single E. coli DnaK. MxDnaK1 is heat-shock inducible, prefers to bind cytosolic proteins, while MxDnaK2 is decreased by heat shock, and is more associated with membrane proteins. The nucleotide binding domain and the substrate binding beta domain are responsible for the significant changes of DnaK substrates, and the former also determines the dimerization of MxDnaK2, but not MxDnaK1. Our work highlights that DnaK is duplicated and regionally evolved for the increase of proteomic complexity in bacteria.

microbiology↗

Immunological findings in a group of individuals who were non-responders to standard two-dose SARS-CoV-2 vaccines

Coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was declared a pandemic. The virus has infected more than 505 million people and caused more than 6 million deaths. However, data on non-responders to SARS-CoV-2 vaccines in the general population are limited. The objective of the study is to comprehensively compare the immunological characteristics of non-responders to SARS-CoV-2 vaccines in the 18-59 years with that in the 60 years and older using internationally recognized cutoff values. Participants included 627 individuals who received physical examinations and volunteered to participate in COVID-19 vaccination from the general population. The main outcome was an effective seroconversion characterized by anti-SARS-CoV-2 spike IgG level of at least 4-fold increase from baseline. Profiling of naive immune cells was analyzed prior to vaccination to demonstrate baseline immunity. Outcomes of effective seroconversion in the 18-59 years with that in the 60 years and older were compared. The quantitative level of the anti-spike IgG was significantly lower in the 60 years and older and in men among the 18-59 years. There were 7.5% of non-responders among the 18-59 years and 11.7% of non-responders in the 60 years and older using the 4-fold increase parameter. The effective seroconversion rate was significantly related to the level of certain immune cells before vaccination, such as CD4 cells, CD8 cells and B cells and the age. An individual with a titer of anti-SARS-CoV-2 spike IgG that is below 50 BAU/mL might be considered a non-responder between 14-90 days after the last vaccine dose. Booster vaccination or additional protective measures should be recommended for non-responders as soon as possible to reduce disease severity and mortality.

immunology↗

ImuA participates in SOS mutagenesis by interacting with RecA1 and ImuB in Myxococcus xanthus

Bacteria have two pathways to restart stalled replication forks caused by environmental stresses, error-prone translesion DNA synthesis (TLS) catalyzed by TLS polymerase and error-free template switching catalyzed by RecA, and their competition on the arrested fork affects bacterial SOS mutagenesis. DnaE2 is an error-prone TLS polymerase, and its functions require ImuA and ImuB. Here we investigated the function of imuA, imuB and dnaE2 in Myxococcus xanthus and found that imuA showed differences from imuB and dnaE2 in bacterial growth, resistance and mutation frequency. Transcriptomics analysis found that ImuA were associated with bacterial SOS response. Yeast-two-hybrid scanning revealed that ImuA interacted with RecA1 besides ImuB. Protein activity analysis proved that ImuA had no DNA binding activity, but inhibited the DNA binding and recombinase activity of RecA1. These findings highlight that ImuA not only participates in TLS by binding ImuB, but also inhibits the recombinase activity of RecA1 in M. xanthus, suggesting a role of ImuA in the two replication restart pathways. ImportanceDnaE2 is responsible for bacterial SOS mutagenesis in nearly one third of sequenced bacterial strains. However, its mechanism, especially the function of its accessory protein ImuA, is still unclear. Here we reported that M. xanthus ImuA might facilitate DnaE2 TLS by inhibiting the recombinase activity of RecA1, which helps to explain the mechanism of DnaE2-dependent TLS and the scientific problem of choosing one of the two restart pathways to repair the stalled replication fork.

microbiology↗

The ParAs function is realized by two separate proteins in the partitioning system of Myxococcus plasmid pMF1

The par operon in the sole myxobacterial plasmid pMF1 includes a function-unknown parC gene in front of the classical parA and parB genes. Removal of parC severely reduced plasmid stability, but ex-situ compensations of parC did not restore the par system function. Individual expression of parA formed insoluble proteins, while co-expression of parC before parA produced a soluble ParC-ParA heterodimer. ParA alone had no ATPase activity and no polymerization, while ParC addition aided ParA to restore the activities. Fusing ParC and ParA in different ways all produced soluble proteins and some restored ATPase activity or increased plasmid stability. Protein interaction model analysis and experiments revealed that ParC structurally mimics the N-terminal of Ia-type SopA (ParA), endowing the Myxococcus ParA protein to play functions by shifting of ParC between two sites on ParA surface. The present results highlight that ParC functions as a part of ParA to support its soluble expression and function, and the separation of ParC and ParA into two proteins in structure enables the ParC fragment to shift in a larger range around ParA to function during partitioning. Author summaryOur work on ParC here provides a new example for the evolution of multi-domain protein. ParC and ParA are two proteins, but their expression and function act as a whole, which proposes a new regulatory model for bacterial par system, and also provides research ideas and materials for the study of functional coordination and evolution of ParA domains in the future.

microbiology↗

Functional divergence and potential mechanisms of the duplicate recA genes in Myxococcus xanthus

RecA is a ubiquitous multifunctional protein for bacterial homologous recombination and SOS response activation. Myxococcus xanthus DK1622 possesses two recA genes, and their functions and mechanisms are almost unclear. Here, we showed that the transcription of recA1 (MXAN_1441) was less than one-tenth of recA2 (MXAN_1388). Expressions of the two recA genes were both induced by ultraviolet (UV) irradiation, but in different periods. Deletion of recA1 did not affect the growth, but significantly decreased the UV-irradiation survival, the homologous recombination ability, and the induction of the LexA-dependent SOS genes. Comparably, the deletion of recA2 markedly prolonged the lag phase for cellular growth and antioxidation of hydrogen peroxide, but did not change the UV-irradiation resistance and the SOS-gene inducibility. The two RecA proteins are both DNA-dependent ATPase enzymes. We demonstrated that RecA1, but not RecA2, had in vitro DNA recombination capacity and LexA-autolysis promotion activity. Transcriptomic analysis indicated that the duplicate RecA2 has evolved to mainly regulate the gene expressions for cellular transportation and antioxidation. We discuss the potential mechanisms for the functional divergence. This is the first time to clearly determine the divergent functions of duplicated recA genes in bacterial cells. The present results highlight that the functional divergence of RecA duplicates facilitates the exertion of multiple RecA functions.\n\nAuthor summaryMyxobacteria has a large-size genome, contains many DNA repeats that are rare in the prokaryotic genome. It encodes bacterial RecA that could promote recombination between homologous DNA sequences. How myxobacteria avoid the undesired recombination between DNA repeats in its genome is an interesting question. M. xanthus encodes two RecA proteins, RecA1 (MXAN_1441) and RecA2 (MXAN_1388). Both RecA1 and RecA2 shows more than 60% sequence consistency with E. coli RecA (EcRecA) and can partly restore the UV resistance of E. coli recA mutant. Here, our results proved their divergent functions of the two RecAs. RecA1 retains the ability to catalyze DNA recombination, but its basal expression level is very low. RecA2 basal expression level is high, but no recombination activity is detected in vitro. This may be a strategy for M. xanthus to adapt to more repetitive sequences in its genome and avoid incorrect recombination.\n\nHighlightsO_LIM. xanthus has two recAs, which are expressed with significantly different levels. Both recAs are inducible by UV irradiation, but in different stages.\nC_LIO_LIThe absence of recA1 reduces bacterial UV-irradiation resistance, while the absence of recA2 delays bacterial growth and antioxidant capacity.\nC_LIO_LIRecA1 retains the DNA recombination and SOS induction abilities, while RecA2 has evolved to regulate the expression of genes for cellular transport and antioxidation.\nC_LI

genetics↗