The epsilon subunit of the ATP synthases from chloroplasts and Escherichia coli regulates the activity of the enzyme and is required for ATP synthesis. The epsilon subunit is not required for the binding of the catalytic portion of the chloroplast ATP synthase (CF1) to the membrane-embedded part (CFo). Thylakoid membranes reconstituted with CF1 lacking its epsilon subunit (CF1-epsilon) have high ATPase activity and no ATP synthesis activity, at least in part because the membranes are very leaky to protons. Either native or recombinant epsilon subunit inhibits ATPase activity and restores low proton permeability and ATP synthesis. In this paper we show that recombinant epsilon subunit from which 45 amino acids were deleted from the C-terminus is as active as full-length epsilon subunit in restoring ATP synthesis to membranes containing CF1-epsilon. However, the truncated form of the epsilon subunit was significantly less effective as an inhibitor of the ATPase activity of CF1-epsilon, both in solution and bound to thylakoid membranes. Thus, the C-terminus of the epsilon subunit is more involved in regulation of activity, by inhibiting ATP hydrolysis, than in ATP synthesis.
<p style="text-align: justify;"><strong>Aims</strong>: The chloroplast DNA sequence of eight Georgian grape cultivars (Rkatsiteli, Saperavi, Meskhuri Mtsvane, Chkhaveri, Aladasturi, Krakhuna, Tsitska, Tsolikouri) and three French cultivars (Chardonnay, Gouais Blanc, Chasselas), belonging to four different haplogroups (AAA, ATT, ATA, GTA), was determined by Illumina resequencing of genomic DNA. The chloroplast DNA sequence of the Maxxa cultivar was used as reference.</p><p style="text-align: justify;"><strong>Methods and results</strong>: The comparison of sequenced chloroplast DNA gave 100 % identity to Chardonnay and Gouais Blanc, differing from Meskhuri Mtsvane by two insertions/deletions (indels) (all ATA haplogroup). The difference between Chasselas and Saperavi was a single insertion (both ATT haplogroup), while Maxxa, Chkhaveri, Aladasturi, Krakhuna, Tsitska and Tsolikouri were all identical (all members of the GTA haplogroup). Forty-seven identical single nucleotide polymorphisms (SNPs) were detected in the AAA, ATA and ATT haplogroups in comparison to the reference DNA. Additionally, 18 SNPs were detected for the ATT haplogroup, 4 for AAA, 6 for ATA and 11 for both AAA and ATA. The phylogenetic results show that the ATT, AAA and ATA haplogroups are more closely related to each other than to the GTA haplogroup.</p><p style="text-align: justify;"><strong>Conclusion</strong>: In the sequencing data of grape genomic DNA at the coverage (read depth) of chromosomal DNA 30-40, the coverage of chloroplast DNA reaches several thousand reads per bp due to the high number of chloroplast DNA copies in genomic DNA, much higher than necessary for resequencing. Based on these data, a new methodology of simultaneous resequencing of large number of chloroplast DNA was developed without preliminary chloroplast isolation or chloroplast enrichment.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: This method has great potential for expanding both phylogenetic and population genetic information on the evolution of domesticated crops.</p>
The main aim of this research was to establish the nucleotide sequence of the highly variable region of the D loop of the mitochondrial DNA of some Georgian domestic animal species (cattle, goat, sheep) as well as their phylogenetic position among the worldwide set of domestic animals. In this study, a total of 5 haplogroups (T – 5; T3 – 7; T1 – 1; T2 – 2; T5 – 2) in 17 Georgian Mountain cattle (GMC), 4 haplogroups (A – 15; A2a1 – 3; A1a – 1; A6 – 3) in 22 Georgian goats and 3 haplogroups (A – 10; B – 16; C -15) in 41 Georgian sheeps (15 Imeretian and 26 Tushetian) were detected. This study represents the first attempt of Genetic study of native Georgian livestocks. The GMC, Georgian (Megrelian) goat, Georgian (Imeretian and Tushetian) sheep mitogenomes were grouped phylogenetically in the haplogroups indicating the closeness to the Near Eastern animals.
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