MOLECULAR-GENETIC ANALYSIS OF REGIONS OF CHROMOSOMEATTACHMENT TO NURSE CELLS NUCLEAR ENVELOPE OF ANOPHELESMALARIA MOSQUITOES FROM «MACULIPENNIS» COMPLEX

Nuclear envelope attachment DNA region from nurse cells XL chromosome ofAnopheles messeae Fall (2b-c region) was isolated by microdissection procedure. ThisDNA was cloned in plasmid vector and sequenced. The analysis in database TBLASTXVectorBase, BLASTN EnsemblMetazoa, FlyBase RepeatMasker, Censor and TandemRepeatsFindersoft reveals that this region consists of transposable elements, tandem repeatsand genes. LTR-retrotransposons, LINE, described in An. gambiae Giles werefound among transposable elements. In 2b-c region transposable elements typical ofother dipterans, protozoa, vertebrates, plants and other organisms were detected. However,MITE SINE and helitrons typical of dipterous were not found in this region. Tandemrepeats are only represented by microsatellites and minisatellites with a small numberof copies. Repeats with AAAAG motifs characteristic for DNA nuclear lamina of micehepatocytes and conservative in evolution were found among tandem repeats. The predominationof repetitive DNA in nuclear envelope attachment region of An. messeae XLchromosome signifies the similarity of this region sequences and constitutive heterochromatin.Similar sequences that characterize nuclear envelope attachment regions of An.messeae XL and 3R chromosomes and An. atroparvus van Thiel. XL chromosome weresearched. Nuclear envelope attachment regions of An. messeae 3R chromosome and An.atroparvus XL chromosome were microdissected. DNA from this regions was used forclone library screening by dot-blot hybridization. Homologous sequences were analyzedwith ChrClass soft to find their potential capacity to bind intranuclear proteins. It wasshown that the major part of these sequences is capable to bind nuclear matrix proteinsincluding nuclear lamina proteins and synaptonemic complex proteins or containSAR/MAR sequences. Twelve clones from An. messeae 2b-c region clones library arehomologous with the sequences from An. messeae 3R chromosome nuclear envelope attachmentregion and eleven clones - with An. atroparvus XL chromosome one. Threeclones were identical to all investigated regions. On the basis of the analysis multiplexnuclear envelope attachment regions organization was supposed possible. Probably, nuclearenvelope attachment regions consist of the following elements: 1) chromosomspecificand evolutionary conservative; 2) specific for every attachment region; 3) universalfor all attachment regions. The obtained results are necessary for estimating DNA sequencessignificance in chromosome architecture reorganization in malaria mosquitoesnurse cells during speciation.

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