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PrO2 is Fluorite structured and crystallizes in the cubic Fm-3m space group. The structure is three-dimensional. Pr4+ is bonded in a body-centered cubic geometry to eight equivalent O2- atoms. All Pr–O bond lengths are 2.48 Å. O2- is bonded to four equivalent Pr4+ atoms to form a mixture of edge and corner-sharing OPr4 tetrahedra.
Mycobacterium smegmatis Nei2 is a monomeric enzyme with AP β-lyase activity on single-stranded DNA. Expression of Nei2, and its operonic neighbor Lhr (a tetrameric 3'-to-5' helicase), is induced in mycobacteria exposed to DNA damaging agents. Here, we find that nei2 deletion sensitizes M. smegmatis to killing by DNA inter-strand crosslinker trimethylpsoralen but not to crosslinkers mitomycin C and cisplatin. By contrast, deletion of lhr sensitizes to killing by all three crosslinking agents. We report a 1.45 Å crystal structure of recombinant Nei2, which is composed of N and C terminal lobes flanking a central groove suitable for DNA binding. The C lobe includes a tetracysteine zinc complex. Mutational analysis identifies the N-terminal proline residue (Pro2 of the ORF) and Lys51, but not Glu3, as essential for AP lyase activity. We find that Nei2 has 5-hydroxyuracil glycosylase activity on single-stranded DNA that is effaced by alanine mutations of Glu3 and Lys51 but not Pro2. Testing complementation of psoralen sensitivity by expression of wild-type and mutant nei2 alleles in Δnei2 cells established that AP lyase activity is neither sufficient nor essential for crosslink repair. By contrast, complementation of psoralen sensitivity of Δlhr cells by mutant lhr alleles depended on Lhr’s ATPase/helicase activities and its tetrameric quaternary structure. The lhr–nei2 operon comprises a unique bacterial system to rectify inter-strand crosslinks.
Most problems considered to be solvable by expert systems have very large search space. It is imperative to use efficient search strategy in expert system tools. Thus, OPS5 uses a kind of hill climbing which is very efficient. However, hill climbing is inadequate for many problems because it is one of the least dependable search strategies. In order to make the search efficient and adequate one can: (1) adopt best-first search instead of hill climbing, or (2) modify hill climbing with intelligent backtracking. The second alternative is adopted. It is implemented in a production system called PRO2 embedded in C running on UNIX. It is called hill tracking. It is a general purpose tool for developing expert systems. This is a rule based production system with an effective, intelligent and flexible backtracking control mechanism, which makes the system more dependable. The advantages and disadvantages of PRO2 are discussed.