Role of the N-terminal region and of β-sheet residue Thr29 on the activity of the ω2 global regulator from the broad-host range Streptococcus pyogenes plasmid pSM19035
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Karin Welfle
Abstract
The dimeric regulatory protein wild-type ω (wt ω2) binds to arrays of 7-bp sequences (heptads) present in the operator DNA region of copy control and partition functions of plasmid pSM19035. Each ω2 protein probably binds with an antiparallel β-sheet structure in the major groove of the 7-bp subsite of the operator DNA. Exchange of threonine at position 29 to alanine (T29A) drastically affects the activity of variant protein ω2T29A both in vivo and in vitro, and reduces the thermodynamic stability ΔGu0, but does not change the conformation. Likewise, the binding affinity to DNA is reduced and the association of the two monomeric subunits of the ω2T29A dimer is weakened, as manifested by an increase in the dissociation constant from 3.2 μM for wt ω2 to 6.3 μM for ω2T29A. Denatured dimers are formed upon thermal unfolding of wt ω2 and ω2T29A at ca. 45 μM (Dn↔Du). Removal of 8 (ω2ΔN8), or even 18 (ω2ΔN18) N-terminal amino acids has no obvious effect either on the core structure or on the activity in comparison to wt ω2. The stability of variants ω2ΔN8 and ω2ΔN18 is similar to that of wt ω2, and their binding to operator DNA is not impaired.
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©2005 by Walter de Gruyter Berlin New York
Articles in the same Issue
- Paper of the Year 2004: Award to Yin Wang and Iris Lorenzi
- Improving the levels of essential amino acids and sulfur metabolites in plants
- Symmetry at the active site of the ribosome: structural and functional implications
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- Developing an effective RNA interference strategy against a plus-strand RNA virus: silencing of coxsackievirus B3 and its cognate coxsackievirus-adenovirus receptor
- NF-κB contributes to transcription of placenta growth factor and interacts with metal responsive transcription factor-1 in hypoxic human cells
- Regulation of asparagine synthetase gene transcription by the basic region leucine zipper transcription factors ATF5 and CHOP
- Role of the N-terminal region and of β-sheet residue Thr29 on the activity of the ω2 global regulator from the broad-host range Streptococcus pyogenes plasmid pSM19035
- Lysine 3 acetylation regulates the phosphorylation of yeast 6-phosphofructo-2-kinase under hypo-osmotic stress
- Determination of the specificity of monoclonal antibodies against Schistosoma mansoni CAA glycoprotein antigen using neoglycoconjugate variants
- Cholesterol is the major component of native lipoproteins activating the p38 mitogen-activated protein kinases
- Focal adhesion kinase is redistributed to focal complexes and mediates cell spreading in macrophages in response to M-CSF
- Inhibition of cathepsin B reduces β-amyloid production in regulated secretory vesicles of neuronal chromaffin cells: evidence for cathepsin B as a candidate β-secretase of Alzheimer's disease
- Activation processing of cathepsin H impairs recognition by its propeptide
Articles in the same Issue
- Paper of the Year 2004: Award to Yin Wang and Iris Lorenzi
- Improving the levels of essential amino acids and sulfur metabolites in plants
- Symmetry at the active site of the ribosome: structural and functional implications
- GPCR-induced migration of breast carcinoma cells depends on both EGFR signal transactivation and EGFR-independent pathways
- Developing an effective RNA interference strategy against a plus-strand RNA virus: silencing of coxsackievirus B3 and its cognate coxsackievirus-adenovirus receptor
- NF-κB contributes to transcription of placenta growth factor and interacts with metal responsive transcription factor-1 in hypoxic human cells
- Regulation of asparagine synthetase gene transcription by the basic region leucine zipper transcription factors ATF5 and CHOP
- Role of the N-terminal region and of β-sheet residue Thr29 on the activity of the ω2 global regulator from the broad-host range Streptococcus pyogenes plasmid pSM19035
- Lysine 3 acetylation regulates the phosphorylation of yeast 6-phosphofructo-2-kinase under hypo-osmotic stress
- Determination of the specificity of monoclonal antibodies against Schistosoma mansoni CAA glycoprotein antigen using neoglycoconjugate variants
- Cholesterol is the major component of native lipoproteins activating the p38 mitogen-activated protein kinases
- Focal adhesion kinase is redistributed to focal complexes and mediates cell spreading in macrophages in response to M-CSF
- Inhibition of cathepsin B reduces β-amyloid production in regulated secretory vesicles of neuronal chromaffin cells: evidence for cathepsin B as a candidate β-secretase of Alzheimer's disease
- Activation processing of cathepsin H impairs recognition by its propeptide
