Protein Interactomes Identify Distinct Pathways forStreptococcus mutansYidC1 and YidC2 Membrane Protein Insertases

Abstract

20Virulence properties of cariogenic Streptococcus mutans depend on integral membrane proteins. 21Bacterial protein trafficking involves the co-translational signal recognition particle (SRP) 22 pathway components Ffh and FtsY, the SecY translocon, and membrane-localized YidC 23 chaperone/insertases. Unlike Escherichia coli, S. mutans survives loss of the SRP pathway. In 24 addition, S. mutans has two yidC paralogs. The yidC2 phenotype largely parallels that of ffh 25 and ftsY while the yidC1 phenotype is less severe. This study defined YidC1 and YidC2 26 interactomes to identify their respective functions alone and in concert with the SRP, ribosome, 27 and/or Sec translocon. A chemical cross-linking approach was employed, whereby whole cell 28 lysates were treated with formaldehyde followed by Western blotting using anti-Ffh, FtsY, 29 YidC1 or YidC2 antibodies and mass spectrometry (MS) analysis of gel-shifted bands. Cross-30 linked lysates from WT and yidC2 strains were also reacted with anti-YidC2 antibodies 31 coupled to magnetic Dynabeads TM , with co-captured proteins identified by MS. Additionally, C-32 terminal tails of YidC1 and YidC2 were engineered as glutathione-S-transferase fusion proteins 33 and subjected to 2D Difference Gel Electrophoresis and MS analysis after being reacted with 34 non-cross-linked lysates. Results indicate that YidC2 works in concert with the SRP-pathway, 35 while YidC1 works in concert with the SecY translocon independently of the SRP. In addition, 36 YidC1 and/or YidC2 can act alone in the insertion of a limited number of small integral 37 membrane proteins. The YidC2-SRP and YidC1/SecY pathways appear to function as part of an 38 integrated machinery that couples translation and transport with cell division, as well as 39 transcription and DNA replication. 40 41 42 Importance 43

Transcriptome Analysis Reveals the Genes Involved in Bifidobacterium Longum FGSZY16M3 Biofilm Formation

“…Moreover, EPS production is often involved in the oxidative stress response [ 61 ]. Almost all gram-positive organisms possess two or more yidC genes [ 33 ]. Three yidC genes (Bl01|peg.766, Bl01|peg.767, and Bl01|peg.1516) were identified in B. longum FGSZY16M3.…”

“…Transcriptome data of B. longum FGSZY16M3 during biofilm formation have been deposited in the National Center for Biotechnology Information Search database (NCBI) under BioProject accession code PRJNA680454. We preprocessed paired-end reads using fastp [ 33 ], which can perform adapter trimming, quality filtering and quality control. Reads were aligned to B. longum FGSZY16M3 genomes using HISAT2 v2.20 [ 34 ].…”

Integration of Transcriptome and Metabolome Reveals the Genes and Metabolites Involved in Bifidobacterium bifidum Biofilm Formation

“…Transcriptome data of B. bifidum FHB150 during biofilm formation have been deposited in the National Center for Biotechnology Information (NCBI) database under BioProject accession code PRJNA733339. The paired-end reads were preprocessed using fastp [ 49 ], and reads were aligned to B. bifidum FHB150 genomes using HISAT2 v2.20 [ 50 ]. Raw read counts were created using featureCounts [ 51 ].…”

Multi-omics analysis reveals genes and metabolites involved in Bifidobacterium pseudocatenulatum biofilm formation

“…Transcriptome data of B. pseudocatenulatum have been stored in the national center for biotechnology information (NCBI), and the BioProject code is PRJNA738670. 1 fastp was used to preprocess paired-end reads (Vasquez et al, 2020). Reads were aligned to B. pseudocatenulatum FJLHD4M2 genomes using HISAT2 v2.20 (Kim et al, 2019).…”