21C1
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 800 bp JEP132 779 bp Composite sequence: CAGNCTGAAT ATCGGTCAGC TGTCGTAGGG CATCAGCTGA AACACCAGGA AAGCAGAGAT CCCCTGTGCC CATATTGAAG ATACTGACAA TATTTATTGA CCTTCTGAAC TGAACAATAG ATTCCCTGTG CGTTGAGAAG ACAATTTCAA GCAGAGCGTC TTCCGCTATG GAAATCAATT CTTCAATCAG TTTTTTAAAA GCTTTTTCAT GCAAAAGCAC ATCCAGTTCG TCAATCAGCA GTAAACCATT AGGCTTCAGA AGGGGGTCAT GAGCGGCTTT CAGCACTTCA AAAACCCGTT TCTCTCCCGC TCCCATGGTA TGTTCAGTGT AGGTAATTCC GTTTTTGGTA AATTTACTGA ATGTGAAGCC ACTGCTGGTC GTACAGGTCA TCAGATCTGA GTAATCCGCT TCAAGCACGC GACACATAGA TTCCAGTATT TTTTGTTTGA TATTGGCGGG TTCGAAGCTT TGGCTTACAT ATTTCGCATG ACGTGAAGCC GCCAGATCGT CGGACAAAGT GCCAAGCGTT TGCAGACCCA GATAGAGCGA CTCCTTAACG GGTCGACGAT CATACACTGG CTGCCAGCGT GCGACTTTAT AATAAGGCTG AGTAAAAGTG TCATTAGGTG CCTGTGTGGG GACAAATAAT ATTTTGTTAC TCTGATTGTT CAGCGTTCCC GCGTAAAAAT GGGCTGTAAA ACCACTGTCA CGCCAGTTGT TATCAACGTG AGGGGTAAAG AAATCAGAGA ACCGGTTATT TTCCTTTGAG GATTATCCCG GGGTTTGTAA CAACAGGCAA AGGCAAGGGC ATGAATAATC GTGGATTTAC CGGAGCCGTT CATCCCCATA ATGGCAGTAA CTTTGCTGTC AGCAGGAAAT GTGATTGTAC AGTTTGAAAT GCCTTTAAGG CTCTGAATCT GTATTTCCTT CAAGCGTAAT TGTGCTGGAA TGTTTTGCCT GGCTGCCATT CTGATCGGTT CCTTTTTATG CTAATAGTCG GATTTATCCC AAGTACACTA CCTTAATTCA GGAAAAAAGT CTGGCATCTA AAGCAGAGAT TTTACAGGCA GATATTACCT ATTCCGATTT TGGGCTTTTT CGGATCTGCG ACGCAAAAAC GCCGACCGCC GCACGGAGCT TTGCTAGCAG ACCAACCAAA TTGTTAGCAA TGGGTCATTG TTGTTATCGC ATTAAAATGA AGCTAGCTGT ATTGGATATA GCATTGTAAT TTTCATTTCT GCTTTATATG GTTTTGCTAT CATCACAATA CATCTCTATC GAAGCAGGAC ACCACTATGC TCAGTCAGAT GAACCTACGC TTTCCGAAAT CCCTGATCGC CGCCCTGAAA AACCGGGCCG CCGCTGAAAC GGTGCCGGTC AACACCCTGG CCGAACGTCT GCTGCAGCTC ACCCAGCATG CCGGAGACGG CTTCCGCCAC CGCGCCTTTC AGGTGGCCAT ACATCTGGCC TTCGAACTCG GCTTCCAGCT GCGCGATGCT CTTGCCGGTC ACACCGGCGA GAATATCCAA CAGGTTGGAG ACGCCCGCCT TGTTGACCAC GTCGTAACGC ACGACAGGC
BLASTX with this sequence (full results here) shows that the gene adjacent to the site of the transposon insertion is homologous to a tryptophanyl-tRNA synthetase.
Taking the other part of the sequence (1-1248; results here), one obtains as best hit:
>gi|281920|pir||S28007 probable ATP-binding protein - Escherichia coli
Length = 492
Score = 40.8 bits (94), Expect = 0.008
Identities = 19/55 (34%), Positives = 33/55 (59%)
Frame = -1
Query: 969 MAARQNIPAQLRLKEIQIQSLKGISNCTITFPADSKVTAIMGMNGSGKSTIIHAL 805
++A+Q QLR+ +I +++ KG + + F T ++G NG GKSTI+ A+
Sbjct: 51 LSAKQLEGGQLRVADIHLENYKGFESLIMDFSMKKNSTILVGNNGCGKSTILDAI 105
The transposon is thus inserted within a protein that contains a domain that is also found in a probable ATP-binding protein. Analysis of the fragment of the putative Serratia protein with Prosite, suggests that it contains an ATP/GTP-binding site motif A (P-loop; GmngsGKS) and perhaps a NACHT-NTPase domain (KVTAIMGMNGSGKSTIIHAL). It therefore is likely to be an ATPase.
If one looks at the E. coli sequence, there is part of a RecF/RecN/SMC N terminal domain:
CD-Length = 170 residues, only 23.5% aligned
Score = 40.6 bits (95), Expect = 1e-04
Query: 66 IHLENYKGF--ESLIMDFSMKKNSTILVGNNGCGKSTILDAI 105 Sbjct: 4 LEIEGFKSYAGKTVIGPFS--PGFNAIVGPNGSGKSNVLDAI 43
Significantly, the similarity stops just at the same position as for the similarity with the putative Serratia sequence.
It is interesting to note that this region is also present in the E. coli proteins that are most closely related to S28007, the probable ATP-binding protein, (see BLASTP results here), and that these are transporters.