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Interactions of archaeal chromatin proteins Alba1 and Alba2 with nucleic acids.

Črnigoj M, Podlesek Z, Zorko M, Jerala R, Anderluh G, Ulrih NP - PLoS ONE (2013)

Bottom Line: Alba2 and equimolar mixtures of Alba1/Alba2 have greater effects on the thermal stability of poly(dA-dT).poly(dA-dT).The secondary structures of the Alba proteins are not significantly influenced by DNA binding, even at high temperatures.Based on these data, we conclude that Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 show different properties in their binding to various DNAs.

View Article: PubMed Central - PubMed

Affiliation: Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.

ABSTRACT

Background: Architectural proteins have important roles in compacting and organising chromosomal DNA. There are two potential histone counterpart peptide sequences (Alba1 and Alba2) in the Aeropyrum pernix genome (APE1832.1 and APE1823).

Methodology/principal findings: THESE TWO PEPTIDES WERE EXPRESSED AND THEIR INTERACTIONS WITH VARIOUS DNAS WERE STUDIED USING A COMBINATION OF VARIOUS EXPERIMENTAL TECHNIQUES: surface plasmon resonance, UV spectrophotometry, circular dichroism-spectropolarimetry, gel-shift assays, and isothermal titration calorimetry.

Conclusions/significance: Our data indicate that there are significant differences in the properties of the Alba1 and Alba2 proteins. Both of these Alba proteins can thermally stabilise DNA polynucleotides, as seen from UV melting curves. Alba2 and equimolar mixtures of Alba1/Alba2 have greater effects on the thermal stability of poly(dA-dT).poly(dA-dT). Surface plasmon resonance sensorgrams for binding of Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 to DNA oligonucleotides show different binding patterns. Circular dichroism indicates that Alba2 has a less-ordered secondary structure than Alba1. The secondary structures of the Alba proteins are not significantly influenced by DNA binding, even at high temperatures. Based on these data, we conclude that Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 show different properties in their binding to various DNAs.

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Surface plasmon resonance sensorgram of the binding of the Alba proteins to the SPRspecAP oligonucleotide.The Alba protein concentration was 6.0 µM. Alba1 (–), Alba2 (·□···□·), the Alba1/Alba2 complex (­­­), and bovine serum albumin (···) at 25°C.
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pone-0058237-g003: Surface plasmon resonance sensorgram of the binding of the Alba proteins to the SPRspecAP oligonucleotide.The Alba protein concentration was 6.0 µM. Alba1 (–), Alba2 (·□···□·), the Alba1/Alba2 complex (­­­), and bovine serum albumin (···) at 25°C.

Mentions: Surface plasmon resonance revealed that both of the Alba proteins bind the DNA oligonucleotides in a complex manner (Figure 3). Bovine serum albumin was used as a negative control, and it showed no DNA binding in all cases. Alba1 showed saturable binding under the conditions used, with fast association. Alba2 showed slower association, and also dissociation, while the sensogram of the Alba1/Alba2 equimolar complex with the SPRspecAP oligonucleotide showed a biphasic pattern of binding to DNA (Figure 3).


Interactions of archaeal chromatin proteins Alba1 and Alba2 with nucleic acids.

Črnigoj M, Podlesek Z, Zorko M, Jerala R, Anderluh G, Ulrih NP - PLoS ONE (2013)

Surface plasmon resonance sensorgram of the binding of the Alba proteins to the SPRspecAP oligonucleotide.The Alba protein concentration was 6.0 µM. Alba1 (–), Alba2 (·□···□·), the Alba1/Alba2 complex (­­­), and bovine serum albumin (···) at 25°C.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585288&req=5

pone-0058237-g003: Surface plasmon resonance sensorgram of the binding of the Alba proteins to the SPRspecAP oligonucleotide.The Alba protein concentration was 6.0 µM. Alba1 (–), Alba2 (·□···□·), the Alba1/Alba2 complex (­­­), and bovine serum albumin (···) at 25°C.
Mentions: Surface plasmon resonance revealed that both of the Alba proteins bind the DNA oligonucleotides in a complex manner (Figure 3). Bovine serum albumin was used as a negative control, and it showed no DNA binding in all cases. Alba1 showed saturable binding under the conditions used, with fast association. Alba2 showed slower association, and also dissociation, while the sensogram of the Alba1/Alba2 equimolar complex with the SPRspecAP oligonucleotide showed a biphasic pattern of binding to DNA (Figure 3).

Bottom Line: Alba2 and equimolar mixtures of Alba1/Alba2 have greater effects on the thermal stability of poly(dA-dT).poly(dA-dT).The secondary structures of the Alba proteins are not significantly influenced by DNA binding, even at high temperatures.Based on these data, we conclude that Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 show different properties in their binding to various DNAs.

View Article: PubMed Central - PubMed

Affiliation: Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.

ABSTRACT

Background: Architectural proteins have important roles in compacting and organising chromosomal DNA. There are two potential histone counterpart peptide sequences (Alba1 and Alba2) in the Aeropyrum pernix genome (APE1832.1 and APE1823).

Methodology/principal findings: THESE TWO PEPTIDES WERE EXPRESSED AND THEIR INTERACTIONS WITH VARIOUS DNAS WERE STUDIED USING A COMBINATION OF VARIOUS EXPERIMENTAL TECHNIQUES: surface plasmon resonance, UV spectrophotometry, circular dichroism-spectropolarimetry, gel-shift assays, and isothermal titration calorimetry.

Conclusions/significance: Our data indicate that there are significant differences in the properties of the Alba1 and Alba2 proteins. Both of these Alba proteins can thermally stabilise DNA polynucleotides, as seen from UV melting curves. Alba2 and equimolar mixtures of Alba1/Alba2 have greater effects on the thermal stability of poly(dA-dT).poly(dA-dT). Surface plasmon resonance sensorgrams for binding of Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 to DNA oligonucleotides show different binding patterns. Circular dichroism indicates that Alba2 has a less-ordered secondary structure than Alba1. The secondary structures of the Alba proteins are not significantly influenced by DNA binding, even at high temperatures. Based on these data, we conclude that Alba1, Alba2, and equimolar mixtures of Alba1/Alba2 show different properties in their binding to various DNAs.

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