A 41–42 bp tandemly repeated sequence isolated from nuclear envelopes of chicken erythrocytes is located predominantly on microchromosomes

Abstract

To study whether specific DNA sequences are associated with nuclear membranes, residual DNA was extracted from DNase-treated nuclear envelopes prepared from erythrocytes of adult chickens (Gallus domesticus). This DNA was then blunt-end ligated into a bacterial plasmid vector. DNA blot analysis and nucleotide sequence determination revealed that approximately 30% of the cloned fragments consisted of different multiples of a 41-42 bp tandemly repeated, partially symmetrical sequence. In situ hybridization to chicken chromosomes demonstrated that the sequence was located primarily on microchromosomes, although some hybridization was also observed to macrochromosomes 7 and 8. Digestion of chicken DNA with any of a number of restriction enzymes did not completely reduce the intensity of a high molecular weight band to which the repeated sequence hybridized. These results, along with those obtained from in situ hybridization, suggested that many copies of this sequence are organized into large tandem arrays, and are not dispersed in many shorter repetitive blocks throughout the chicken genome. Although the repetitive sequence constituted approximately 10% of the chicken genome, it did not hybridize to quail or turkey DNA.

Centromere positions in chicken and Japanese quail chromosomes: de novo centromere formation versus pericentric inversions

“…For a long time, the information about centromerespecific sequences in birds was limited to a few satellites identified in several species (Matzke et al 1990;Solovei et al 1996;Tanaka et al 2000;Saifitdinova et al 2001;Wang et al 2002;Krasikova et al 2006;Deryusheva et al 2007). Even in extensively studied galliform species, such as chicken and quail, primarily microchromosome-specific centromeric repeats were well characterized (Matzke et al 1990;Tanaka et al 2000;Krasikova et al 2006;Deryusheva et al 2007).…”

“…Even in extensively studied galliform species, such as chicken and quail, primarily microchromosome-specific centromeric repeats were well characterized (Matzke et al 1990;Tanaka et al 2000;Krasikova et al 2006;Deryusheva et al 2007). A recent comprehensive analysis revealed elusive centromere DNA specific for chicken chromosomes (Shang et al 2010).…”

“…DNA of BAC clones and chromosome paints were labeled with biotin-dUTP or digoxigenin-dUTP in a DOP-PCR using 6MW primer (Telenius et al 1992). Oligonucleotide probes specific for chicken CNM (Matzke et al 1990) and quail BglII-repeat (Tanaka et al 2000) were also used for centromere detection, namely, CNMpos, CNMneg, and CCOneg oligonucleotides (Deryusheva et al 2007) labeled with Cy3 or Cy5. The labeled probes were dissolved in a standard formamide hybridization buffer with an excess of salmon sperm DNA as described elsewhere Krasikova et al 2006;Deryusheva et al 2007).…”

High-resolution mapping and transcriptional activity analysis of chicken centromere sequences on giant lampbrush chromosomes

“…Chicken CNM repeat (Matzke et al 1990; NCBI accession number X51431) and Japanese quail BglIIrepeat (Tanaka et al 2000; NCBI accession numbers AB035968-AB035970) were mapped to lampbrush chromosomes by FISH with specific oligonucleotide probes as described previously (Krasikova et al 2006;Deryusheva et al 2007). …”

“…For example, deletion of CenZ centromere DNA led to non-appearance of a kinetochore on chromosome Z in chicken DT40 cells (Shang et al 2010). Other macrochromosomes, namely GGA6 and GGA9, and most of the gene-rich microchromosomes, typical for Galliform karyotypes, contain an identical 41 bp-repeat (CNM repeat) within their centromere regions (Matzke et al 1990). CNM repeat is not centromere-specific and in addition to centromere regions localises in subtelomeric and interstitial regions of certain chicken chromosomes (Krasikova et al 2006).…”