Abstract:
:The promoter -10 binding region of the Bacillus subtilis sigma(A) factor forms an amphiphilic alpha-helix with three conserved isoleucines located at four-residue intervals. To investigate the structural and functional roles of the three isoleucine residues, we constructed a series of sigA mutants with single and double Ile-to-Ala substitutions on the hydrophobic face of this alpha-helix and isolated intragenic revertants with either same-site or second-site suppressor that partially restores the structural stability and transcription activity of the mutant sigma(A) factors. Our data show that the three conserved isoleucine residues (Ile-194, Ile-198, and Ile-202) are involved in the hydrophobic core packing of sigma(A); they affect differentially and additively the structure and function of sigma(A), with the central isoleucine residue (Ile-198) playing the most important role. By analogy with the crystal structure of a sigma(70) peptide, it is apparent that interdigital interactions exist between the three conserved isoleucine residues and certain hydrophobic amino acids in region 2. 1 of sigma(A). They include at least the van der Waals contacts between Ile-194 and both Leu-145 and Ile-149, between Ile-198 and both Ile-149 and Tyr-153, as well as between Ile-202 and Tyr-153. The same-site suppressors, Val-194 and Val-198, restore the structural stability and transcription activity of sigma(A) by repacking the hydrophobic core of sigma(A). The second-site suppressor (S291F) appears to be allele-specific, but it is not as effective as the same-site suppressors in restoring sigma(A) structure and function.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Liao CT,Wang WH,Yang HS,Chen JP,Chang BYdoi
10.1093/oxfordjournals.jbchem.a022474keywords:
subject
Has Abstractpub_date
1999-09-01 00:00:00pages
461-9issue
3eissn
0021-924Xissn
1756-2651journal_volume
126pub_type
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journal_title:Journal of biochemistry
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pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a133504
更新日期:1981-08-01 00:00:00
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journal_title:Journal of biochemistry
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1991-07-01 00:00:00
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更新日期:1983-12-01 00:00:00
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更新日期:1995-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:1981-03-01 00:00:00