8E2
Site of transposon insertion cloned as PstI fragment into pSP72. Sequenced in opposite orientations with JEP131 and JEP132, from sites at opposing ends of the transposon, to give: JEP131 580 bp JEP132 582 bp Composite sequence: TTCNCGNNCA TGAGNGCGTC GGCAACCGTT GANCGCCTGC TACACGTGAA GCCTGCGCNC NCGCTANCCG AGGCTTCTTG GGCCGCCACG TGACNAAGGC AGCAGAATNC GAATACGGCT CTTCGCTCTC CGAGCGCCAN CAGTTCGTCG GCGAGTTCAC CGGCGTGCGC CGCGAAGGCT GGGCGGAAGT CGACGTGAAG AATAAATTCT CACTCGGCGA CAGCGTCGAG ATGATGACGC CAGGCGGCAA CGTGCTGTTC ACCCTCGAAA GCATGCAAAA TAAGAAAGGC GAGCCGATTG AGGTCGCGCC GGGCAACGGC CATATCGTTT ATTTGCCGAT CCCGCAGGAT ATTGATCTCA ATTATGCATT GCTGATCCGC AATTTGGCGG AAGAAAATAG CGCCGGAGCC TAACAATCAA TCGTCGCCAT TCGGCGGCGA TTTTTAGCAA ATATTAGAAT TGGATCACAT CAATGTGCCC GCAGCTTGGT TACTATCACG CTGCTTAAAA CGAACAACGA AAATATTTGC ATAGCAATAC TAGGAACCAC CTCCTTAGCC GGCCCATNTA GGTNGCAAAA TTGNGAACCC TTGGGCTGGC TTTTTCTTTA TGGGTCCGCC ATTGCCGCCG GCGCGCCTAT TTTTCATTAC GTTTTCCCGT TATCCATTTT AATAAAAATA ATAACGCCAC ATCGATATAT AAATAACGCT ATAGCGATTA ATCCTGCCGC CATTAAAATT CCGCTTATTG TCAGGTTAAA TTCAGATAAA TTCGCATCGT CTTCATATTC ACTGGCACAA TAGGAAATAA CCGGAATTAG CTGAATATTC TGATGGCTTG GCGGCGTAGG CTGGTTTATC TTGAGGCATT CTTGGTTTTC CGCCGGAGGA GATCGGTAAT GCCTCAACAC TCACATGCAG CCGCCCTGCT TATTCTCAAC GGCAAGGGCG CCGGCAACGA AGAACTGCGT CAGGCGGCAC ACGGCTGCGC GAAGAGCAAA TACGCTGTAC GTGCGCGTGA CCTGGGAGCA CGGNGACGCC GCGCGCTACG TCGCGGAAGC TGACACTTGG CGTCGGGANC CGTCGTCCCG CGGCGGNGCG GACCATAACG AAGNGGCCGC CGGCTGGGCA GCTGCGGNNA NACCGGCGNN TGGGATNTGC G
BLASTX with this sequence hits two different families of proteins (full results here), with the top match for each being: >gi|16123048|ref|NP_406361.1| (NC_003143) putative protease [Yersinia pestis] Length = 464 Score = 158 bits (399), Expect = 3e-38 Identities = 79/111 (71%), Positives = 92/111 (82%), Gaps = 1/111 (0%) Frame = +3 Query: 72 GFLGRHVTKAAEX-EYGSSLSERXQFVGEFTGVRREGWAEVDVKNKFSLGDSVEMMTPGG 248 GFL RHV + + EYG S+SER QFVGEFTGVRREG AEV VKNKFS GDSVE+MTP G Sbjct: 347 GFLRRHVHEEQQTYEYGYSVSERQQFVGEFTGVRREGLAEVLVKNKFSRGDSVELMTPKG 406 Query: 249 NVLFTLESMQNKKGEPIEVAPGNGHIVYLPIPQDIDLNYALLIRNLAEENS 401 N+ FTLESMQ+KKG+P ++APGNGHI+YLPIP++ID+ Y LLIRNL NS Sbjct: 407 NIQFTLESMQDKKGQPADIAPGNGHIMYLPIPEEIDVEYGLLIRNLNGTNS 457 >gi|16123049|ref|NP_406362.1| (NC_003143) putative diacylglycerol kinase [Yersinia pestis] Length = 296 Score = 43.1 bits (100), Expect(2) = 5e-06 Identities = 20/34 (58%), Positives = 22/34 (63%) Frame = +3 Query: 1002 TLYVRVTWEHGDAARYVAEADTWRRXPSSRGGAD 1103 TL+VR+TWEHGDA RYV EA T GG D Sbjct: 31 TLHVRITWEHGDAKRYVEEAATLAVSTVIAGGGD 64 Score = 27.7 bits (60), Expect(2) = 5e-06 Identities = 12/17 (70%), Positives = 15/17 (87%) Frame = +2 Query: 926 LLILNGKGAGNEELRQA 976 LLILNGK +GN E+R+A Sbjct: 5 LLILNGKESGNPEVREA 21
The transposon is thus inserted between two genes. It is downstream of a homologue of a putative protease (YPO2855), and upstream of a homologue of a putative diacylglycerol kinase (YPO2856).
The E. coli homologue of YPO2856, b2086, corresponds to yegS, a gene of unknown function, annotated in Ecogene as being flanked by genes of unknown function (see also the Colibri entry). b2086 is somewhat similar (~30% identity) to bmrU, a protein of unknown function that might be involved in stress responses as, "Transcription of bmrU was very low during exponential growth but increased severalfold after heat stress, exposure to ethanol and salt as well as after glucose starvation", (see Petersohn et al. 1999). The E. coli homologue of YPO2855 is yegQ.
As can be deduced from their names, in Y. pestis YPO2855 and YPO2856 are direct neighbours, while in E. coli, yegQ and yegS are separated by yegR (see Ecogene; note that the Colibri annotation in this region is different).
BLASTP analysis with YPO2856 suggests that there is no strong reason to believe that it is a diacylglycerol kinase. On the otherhand, its homologues are found associated with metabolic enzymes. For example, the Pseudomonas homologue of YPO2856, PA3023, is next to two genes involved in aerobic sugar metabolism. Interestingly, in Pseudomonas, the homologue of YPO2855, PA5440, is far from PA3023, but next to a probable glucose-6-phosphate dehydrogenase. As chance would have it, it is also within 20 kb of wzm (see the entry for clone 20C2).
STRING analysis of the protease (see colour code) and the so-called diacylglycerol kinase (see colour code) does not suggests that they occur close together in other species.
And according to the Enteric server, yegQ and yegS/b2086 are separated by a region that is not conserved between bacteria (see results here or download the PDF file).
It is, however, interesting to see that the pair of genes immediately upstream of yeqS/b2086, are baeS and baeR (see entry for clone 8G1).
While not expected to be affected by the transposon insertion, it is interesting to note that the homologue of the upstream gene YPO2855 contains a U32 peptidase domain, and is very similar to the E. coli protein yegQ (see BLASTP results). These proteins are related to the Proteus mirabilis STM-proteaseA, a known virulence factor, implicated in urinary tract infectivity (reviewed by Coker et al., 2000).
