Development-dependent changes in the tight DNA-protein complexes of barley on chromosome and gene level

Abstract

Background: The tightly bound to DNA proteins (TBPs) is a protein group that remains attached to DNA with covalent or non-covalent bonds after its deproteinisation. The functional role of this group is as yet not completely understood. The main goal of this study was to evaluate tissue specific changes in the TBP distribution in barley genes and chromosomes in different phases of shoot and seed development. We have: 1. investigated the TBP distribution along Amy32b and Bmy1 genes encoding low pI α-amylase A and endosperm specific β-amylase correspondingly using oligonucleotide DNA arrays; 2. characterized the polypeptide spectrum of TBP and proteins with affinity to TBP-associated DNA; 3. localized the distribution of DNA complexes with TBP (TBP-DNA) on barley 1H and 7H chromosomes using mapped markers; 4. compared the chromosomal distribution of TBP-DNA complexes to the distribution of the nuclear matrix attachment sites.

Tightly bound to DNA proteins: Possible universal substrates for intranuclear processes

“…The recently performed experiments on characterization of the TBPs distribution at the gene and chromosome levels by means of hybridization with microarrays of genomic probes and PCR confirm the above conclusion (Bielskiene et al, 2008;Sjakste et al, 2005Sjakste et al, , 2009). …”

“…The use of RNase and restriction endonucleases was also excluded from the protocol of TBPs preparation because purified enzymes added in high concentrations to isolated DNA form stable artifact DNA-protein complexes (Benore-Parsons and Ayoub, 1997). It appeared that the TBPs set is different in barley seedling leaves, roots, and coleoptiles Sjakste et al, 2009). In some cases it is also possible to reveal changes in the TBPs spectrum during seedling development .…”

“…Despite promising results, these studies were abandoned mostly for subjective reasons. Recently we have resumed research on TBPs applying modern research methods (Bielskiene et al, 2008;Sjakste et al, 2005Sjakste et al, , 2009). In the present review we will summarize the formerly published and recent data on TBPs and analyze a possible role of this protein group in intranuclear processes.…”

<i>Hordeum vulgare</i> miežių lapų stabilių DNR baltymų kompleksų baltymų sudėties analizė

“…Despite a great deal of research, the functional significance of TBP is not yet clear. It has been proposed that TBP proteins may participate in differentiation and development processes [4]. Investigation of tightly bound DNA-protein complexes from different barley organs (coleoptile, leaf and roots) from Zadoks stages 07 and 10 can reveal some aspects of this assumption.…”

“…TBP have been found in DNA of numerous evolutionary distant species [3][4][5]. Some of these TBP proteins have been characterized in Ehrlich ascites [6-8] and yeast (Saccharomyces cerevisiae) [9] cells.…”