The Sso7d DNA‐binding protein from <i>Sulfolobus solfataricus</i> has ribonuclease activity

Shehi, Erlet; Serina, Stefania; Fumagalli, Gabriele; Vanoni, Marco; Consonni, Roberto; Zetta, Lucia; Dehò, Gianni; Tortora, Paolo; Fusi, Paola

doi:10.1016/s0014-5793(01)02455-3

Cited by 25 publications

(16 citation statements)

References 34 publications

(56 reference statements)

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“…For this reason, we further investigated the effect of the protonation state of residue Glu35 on the surface potential of the protein. Most interestingly, mutants E35Q and E35L were almost completely inactive and completely inactive, respectively (22).…”

Section: Resultsmentioning

confidence: 99%

Investigations of Sso7d Catalytic Residues by NMR Titration Shifts and Electrostatic Calculations

et al. 2003

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Sso7d is a small basic protein consisting of 62 amino acids isolated from the thermoacidophilic archeobacterium Sulfolobus solfataricus. The protein is endowed with DNA binding properties, RNase activity, and the capability of rescuing aggregated proteins in the presence of ATP. In this study, the electrostatic properties of Sso7d are investigated by using the Poisson-Boltzmann calculation of the surface potential distribution and following by NMR spectroscopy the proton chemical shift pH titration of acidic residues. Although the details of the catalytic mechanism still have to be defined, the results from NMR experiments confirm the possible involvement of Glu35 as the proton acceptor in the catalytic reaction, as seen by its abnormally high pK(a) value. Poisson-Boltzmann calculations and NMR titration shifts suggest the presence of a possible hydrogen bond between Glu35 and Tyr33, with a consequent rather rigid arrangement at these positions. Comparison with RNase T1 suggests that Tyr7 may be a good candidate for acting as a proton donor in the active site of Sso7d as shown by its low phenolic pK(a) of approximately 9.3. Titration experiments performed with the UpA, a RNA dinucleotide model, showed that the protein residues affected by the interaction are mainly located in a different region with respect to the surface affected by DNA recognition, in good agreement with the surface potential distribution found with electrostatic calculations.

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Section: Resultsmentioning

confidence: 99%

Investigations of Sso7d Catalytic Residues by NMR Titration Shifts and Electrostatic Calculations

et al. 2003

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“…Interestingly, single point mutations K12L and E35L completely abolish the RNase activity of Sso7d without affecting its thermal stability. 22 Therefore, an E35L mutant of Sso7d was used as the starting point for construction of the library. The DNA sequences encoding the Sso7d mutant library were obtained through synthesis of a single oligonucleotide with NNN degenerate codons at positions encoding the residues K20, K21, W23, V25, M28, S30, T32, T40, R42 and A44.…”

Section: Resultsmentioning

confidence: 99%

Highly Stable Binding Proteins Derived from the Hyperthermophilic Sso7d Scaffold

Gera

Hussain

Wright

et al. 2011

Journal of Molecular Biology

150

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“…It is not possible to mention detailed here all; Sso7d protein having RNase activity from Sulfolobus solfataricus (Shehi et al 2001), RNase R from Haloferax volcanii (Portnoy and Schuster 2006) and RNase J from Methanocaldococcus jannaschii (Levy et al 2011) can be given as examples for archaeal RNases. Extremophile members of Archaea can provide novel RNases with different characteristics offering promising potential for application in different fields.…”

Section: Production Of Bacterial and Archaeal Rnasesmentioning

confidence: 99%

Microbial ribonucleases (RNases): production and application potential

Hameş

Demir

2015

World J Microbiol Biotechnol

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Ribonuclease (RNase) is hydrolytic enzyme that catalyzes the cleavage of phosphodiester bonds in RNA. RNases play an important role in the metabolism of cellular RNAs, such as mRNA and rRNA or tRNA maturation. Besides their cellular roles, RNases possess biological activity, cell stimulating properties, cytotoxicity and genotoxicity. Cytotoxic effect of particular microbial RNases was comparable to that of animal derived counterparts. In this respect, microbial RNases have a therapeutic potential as anti-tumor drugs. The significant development of DNA vaccines and the progress of gene therapy trials increased the need for RNases in downstream processes. In addition, RNases are used in different fields, such as food industry for single cell protein preparations, and in some molecular biological studies for the synthesis of specific nucleotides, identifying RNA metabolism and the relationship between protein structure and function. In some cases, the use of bovine or other animal-derived RNases have increased the difficulties due to the safety and regulatory issues. Microbial RNases have promising potential mainly for pharmaceutical purposes as well as downstream processing. Therefore, an effort has been given to determination of optimum fermentation conditions to maximize RNase production from different bacterial and fungal producers. Also immobilization or strain development experiments have been carried out.

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The Sso7d DNA‐binding protein from Sulfolobus solfataricus has ribonuclease activity

Cited by 25 publications

References 34 publications

Investigations of Sso7d Catalytic Residues by NMR Titration Shifts and Electrostatic Calculations

Investigations of Sso7d Catalytic Residues by NMR Titration Shifts and Electrostatic Calculations

Highly Stable Binding Proteins Derived from the Hyperthermophilic Sso7d Scaffold

Microbial ribonucleases (RNases): production and application potential

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