, 2009). A close inspection of the PhaR-binding sequences of these genes

revealed a very striking similarity among them. The PhaR-binding sequence present in the phaZ promoter is CTGCCATGCAG (located at nucleotides −77 to −67 relative to the initiation codon of phaZ). The one located in the phaC promoter is CTGCATGGCAG (nucleotides −35 to −25 relative to the initiation codon of phaC), and that in the phaR promoter is CTGCAGCCGCAG (located at nucleotides −31 to −20 relative to the initiation codon of phaR). The only difference among these sequences is in the space Proteases inhibitor region. The sequence of the spacer region of the PhaR-binding motif of the phaZ gene is CAT, and those for phaC and phaR are ATG and AGCC, respectively. This is consistent with our finding that the sequences in the two dyad regions of the PhaR-binding motif cannot be changed, but that in the spacer can be substituted by any three or four nucleotides (Fig. 2). Although PhaR can bind to the promoter regions of phaP, phaR, phaZ, and phaC, it regulates these genes differently. PhaR represses the expression of phaP, HSP inhibitor drugs phaR, and phaZ, but not phaC (Chou et al., 2009). The phaZ and phaC genes are located next to each other in an opposite direction and share the same PhaR-binding motif.

However, binding of PhaR to this motif inhibits phaZ expression, but has no suppressive effect on phaC (Chou et al., 2009). This interesting regulatory mechanism is being investigated. We thank Chao-Hung Lee for valuable discussions and critical editing of the manuscript. This study was supported by a grant (NSC96-2311-B-030-001) from the National Science Council, Arachidonate 15-lipoxygenase Taipei, Taiwan. “
“Alarmone Guanosine 5′-diphosphate (or 5′-triphosphate) 3′-diphosphate [(p)ppGpp]

is the key component that globally regulates stringent control in bacteria. There are two homologous enzymes, RelA and SpoT in Escherichia coli, which are responsible for fluctuations in (p)ppGpp concentration inside the cell, whereas there exists only a single RelA/SpoT enzyme in Gram-positive bacteria. We have identified a bifunctional enzyme with (p)ppGpp-hydrolase/synthase activity in Leptospira interrogans. We show that the relLin gene (LA_3085) encodes a protein that fully complements the relA/spoT double mutants in E. coli. The protein functions as a (p)ppGpp degradase as well as a (p)ppGpp synthase when the cells encounter amino acid stress and deprivation of carbon sources. N-terminus HD and RSD domains of relLin (relLinN) were observed to restore growth of double mutants of E. coli. Finally, We demonstrate that purified RelLin and RelLinN show high (p)ppGpp synthesis activity in vitro. Taken together, our results suggest that L. interrogans contain a single Rel-like bifunctional protein, RelLin, which plays an important role in maintaining the basal level of (p)ppGpp in the cell potentially contributing to the regulation of bacterial stress response.