Article Figures & Data
Figures
- FIG. 1.
Homology of Longus and CFA/III gene clusters. Arrows indicate the direction of transcription. Genes in black correspond to major structural subunit genes.
- FIG. 2.
Maximum-likelihood phylograms of Longus sequences. (a) Unrooted phylogram of lngA sequences. lngA sequences from 21 ETEC strains along with cofA sequences from CFA/III ETEC strains were included. (b) Phylogram of concatenated nonstructural genes in the Longus (based on a subset of six strains) and CFA/III gene clusters. The intra-cof (260-1/E2528C/M403-C1) comparison of nonstructural genes is based on 4 out of 13 loci (cofC, -E, -H, and -P). (c) Phylogram of concatenated adk and fumC genes from 21 ETEC strains. The branch lengths refer to numbers of nucleotide differences. For panels A and B, two different length scales are used. The symbol # indicates strains in which the non-lngA genes were sequenced (see panel B). Homologous lngA allelic groups are denoted by underlined (group 1), bold (group 2), and plain (group 3) text. Strains from which sequences were derived are represented by three-letter designations. lngA group 1 strains: E9034A, L1A; 01117-5, L1B; B2C, L1C; M104, L1D; M145-C2, L1E; M424-C1, L1F; M526-C6B, L1G; P307, L1H; M408-C1, L1I. lngA group 2 strains: 10159-a, L2A; 11381a, L2B, 2108-2, L2C; B7A, L2D; G1026, L2E; M445-C1, L2F; M452-C1, L2G; M626-C, L2H; M633-C1, L2I; MP215-1, L2J; BR5, L2K. lngA group 3 strain ECOR27, L3A. cofA group 1 strains: 2528C, C1A; 260-1, C1B. cofA group 2 strain M403-C1, C2A.
Tables
- TABLE 1.
Host strains and plasmids used or generated in this study
Strain or plasmid Description Reference or source pE9034A ETEC strain E9034A virulence plasmid; carries Longus, CS3, ST, and LT genes 13 E. coli DH5α supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 Laboratory collection E. coli HB101 supE44 Δ(mcrC-mrr) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 leuB6 thi-1 Laboratory collection pBluescript SKII Cloning vector (Apr) Stratagene, La Jolla, CA pCR2.1 Cloning vector (Apr Kmr) Invitrogen, Carlsbad, CA pACYC184 Cloning vector (Cmr Tetr) New England BioLabs, Ipswich, MA pJAG1 5.0-kb BamHI fragment containing Longus genes cloned into pACYC177 11 pOG140 pBluescript SK with 1.0-kb EcoRV insert containing entire lngA gene 13 pOG341 pACYC184 with 8-kb SalI insert containing lngF to lngP This study pOG347 pACYC184 with 5-kb EcoRI insert containing lngR to lngA This study pOG349 pACYC184 with 3-kb EcoRI insert containing lngG to lngH This study pOG378 pACYC184 with 3-kb EcoRI insert containing lngT to lngB This study pOG380 pACYC184 with 5-kb NcoI insert containing lngJ to lngP This study pOG381 Same as pOG380 but with inverted 5-kb NcoI insert containing lngJ to lngP This study pOG371 pCR2.1 with 1.5-kb lngR-lngS insert This study - TABLE 2.
Biosynthetic Longus genes, putative functions, gene homologues, and allelic variations
Gene Length (bp) % Identity to CFA/III gene/protein Putative function Type IV pilus gene homologs Dn/Dsd of allelic variants lngX1 378 NAa Unknown None NA lngR 303 72/72 Gene regulation cofR, clpB, pefB; PapB regulator family 0.011/0.010 lngS 894 68/70 Gene regulation cofS, perA, toxT; AraC regulator family 0.004/0.015 lngT 441 79/82 Transglycosylase cofT, bfpH 0.005/0.012 lngX2 219 NA Unknown None NA lngA 708 74/79 Structural subunit cofA, cfcA, bfpA, tcpA 0.053/0.571 lngB 1,572 74/78 Pilin-like subunit cofB, cfcB, tcpB 0.005/0.068 lngC 411 7370 Unknown cofC, cfcC, tcpQ, bfpG 0.001/0.016 lngD 1,476 81/91 Outer membrane lipoprotein cofD, cfcD, bfpB, tcpC, pulD 0.001/0.010 lngE 558 75/75 Inner membrane cofE, cfcE, tcpR 0.003/0.010 lngF 771 68/66 Inner membrane cofF, cfcF, tcpD 0.004/0.014 lngG 483 73/75 Periplasmic protein cofG, cfcG, bfpU, tcpS 0.003/0.016 lngH 1,332 73/75 Nucleotide binding, twitching motility cofH, cfcH, bfpD, tcpT, pulE 0.001/0.016 lngI 969 83/96 Inner membrane cofI, cfcI, bfpE, tcpE, pulF 0.002/0.026 lngJ 1,119 63/57 ATPase, twitching motility cofJ, cfcJ, bfpF, tcpF 0.002/0.033 lngP 852 62/59 Prepilin peptidase cofP, cfcP, bfpP, tcpJ, pulO 0.010/0.038 adk b 534c 0.0005/0.025 fumC b 465c 0.004/0.072
Supplemental material
Files in this Data Supplement:
- Supplemental file 1
-
Fig. S1, distribution of polymorphic sites across LngA and CofA variants
Zipped PowerPoint document, 21K. - Supplemental file 2
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Table S1, ETEC isolates screened for longus and CFA/III by PCR
MS Word document, 76K.
- Supplemental file 1
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Fig. S1, distribution of polymorphic sites across LngA and CofA variants
