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Addition of an acetyl group, which carries a negative charge, effectively removes the positive charge and hence, reduces the interaction between the histone tail and the nucleosome. [18] This opens up the usually tightly packed nucleosome and allows transcription machinery to come into contact with the DNA template, leading to gene transcription.
Core histones are four proteins called H2A, H2B, H3 and H4 and they are all found in equal parts in the cell. All four of the core histone amino acid sequences contain between 20 and 24% of lysine and arginine and the size or the protein ranges between 11400 and 15400 daltons, making them relatively small, yet highly positively charged proteins. [6]
The negative charge of its phosphate backbone moves the DNA towards the positively charged anode during electrophoresis. However, the migration of DNA molecules in solution, in the absence of a gel matrix, is independent of molecular weight during electrophoresis, i.e. there is no separation by size without a gel matrix. [12]
DNA gel electrophoresis is usually performed for analytical purposes, often after amplification of DNA via polymerase chain reaction (PCR), but may be used as a preparative technique prior to use of other methods such as mass spectrometry, RFLP, PCR, cloning, DNA sequencing, or southern blotting for further characterization.
For example, the positive charge of ethidium bromide can reduce the DNA movement by 15%. [12] Agarose gel electrophoresis can be used to resolve circular DNA with different supercoiling topology. [16] DNA damage due to increased cross-linking will also reduce electrophoretic DNA migration in a dose-dependent way. [17] [18]
Histone acetyltransferases transfer an acetyl group to specific lysine residues on histones, which neutralizes their positive charge and thus reduces the strong interactions between the histone and DNA. [25] Acetylation is also thought to perturb interactions between individual nucleosomes and act as interaction sites for other DNA-associated ...
DNA exists in many possible conformations that include A-DNA, B-DNA, and Z-DNA forms, although only B-DNA and Z-DNA have been directly observed in functional organisms. [14] The conformation that DNA adopts depends on the hydration level, DNA sequence, the amount and direction of supercoiling, chemical modifications of the bases, the type and ...
In eukaryotic cells, DNA is associated with about an equal mass of histone proteins in a highly condensed nucleoprotein complex called chromatin. [14] Deoxyribonucleoproteins in this kind of complex interact to generate a multiprotein regulatory complex in which the intervening DNA is looped or wound.