title: Intermediate introns in nuclear genes of euglenids - are they a distinct type?
abstract: BACKGROUND
  Nuclear genes of euglenids contain two major types of introns: conventional spliceosomal and nonconventional introns. The latter are characterized by variable non-canonical borders, RNA secondary structure that brings intron ends together, and an unknown mechanism of removal. Some researchers also distinguish intermediate introns, which combine features of both types. They form a stable RNA secondary structure and are classified into two subtypes depending on whether they contain one (intermediate/nonconventional subtype) or both (conventional/intermediate subtype) canonical spliceosomal borders. However, it has been also postulated that most introns classified as intermediate could simply be special cases of conventional or nonconventional introns.
  
  
  RESULTS
  Sequences of tubB, hsp90 and gapC genes from six strains of Euglena agilis were obtained. They contain four, six, and two or three introns, respectively (the third intron in the gapC gene is unique for just one strain). Conventional introns were present at three positions: two in the tubB gene (at one position conventional/intermediate introns were also found) and one in the gapC gene. Nonconventional introns are present at ten positions: two in the tubB gene (at one position intermediate/nonconventional introns were also found), six in hsp90 (at four positions intermediate/nonconventional introns were also found), and two in the gapC gene.
  
  
  CONCLUSIONS
  Sequence and RNA secondary structure analyses of nonconventional introns confirmed that their most strongly conserved elements are base pairing nucleotides at positions +4, +5 and +6/ -8, -7 and -6 (in most introns CAG/CTG nucleotides were observed). It was also confirmed that the presence of the 5' GT/C end in intermediate/nonconventional introns is not the result of kinship with conventional introns, but is due to evolutionary pressure to preserve the purine at the 5' end. However, an example of a nonconventional intron with GC-AG ends was shown, suggesting the possibility of intron type conversion between nonconventional and conventional. Furthermore, an analysis of conventional introns revealed that the ability to form a stable RNA secondary structure by some introns is probably not a result of their relationship with nonconventional introns. It was also shown that acquisition of new nonconventional introns is an ongoing process and can be observed at the level of a single species. In the recently acquired intron in the gapC gene an extended direct repeats at the intron-exon junctions are present, suggesting that double-strand break repair process could be the source of new nonconventional introns.
authors.affiliation: Department of Molecular Phylogenetics and Evolution, Institute of Botany, Faculty of Biology, University of Warsaw, Warsaw, Poland. milan@biol.uw.edu.pl.
authors.forename: Rafał
authors.initials: R
authors.lastname: Milanowski
authors.affiliation: Department of Molecular Phylogenetics and Evolution, Institute of Botany, Faculty of Biology, University of Warsaw, Warsaw, Poland. n.guminska@biol.uw.edu.pl.
authors.forename: Natalia
authors.initials: N
authors.lastname: Gumińska
authors.affiliation: Department of Molecular Phylogenetics and Evolution, Institute of Botany, Faculty of Biology, University of Warsaw, Warsaw, Poland. ankarn@biol.uw.edu.pl.|Department of Botany, University of British Columbia, Vancouver, Canada. ankarn@biol.uw.edu.pl.
authors.forename: Anna
authors.initials: A
authors.lastname: Karnkowska
authors.affiliation: Department of Molecular Biology, Institute of Biochemistry, Faculty of Biology, University of Warsaw, Warsaw, Poland. takao@biol.uw.edu.pl.
authors.forename: Takao
authors.initials: T
authors.lastname: Ishikawa
authors.affiliation: Department of Molecular Phylogenetics and Evolution, Institute of Botany, Faculty of Biology, University of Warsaw, Warsaw, Poland. zakrys@biol.uw.edu.pl.
authors.forename: Bożena
authors.initials: B
authors.lastname: Zakryś
mesh_terms: D020029:Base Pairing; D001483:Base Sequence; D016822:Euglenida / Q000235:genetics*; D005091:Exons; D017125:Genes, Protozoan; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics; D007438:Introns; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000235:genetics; D017422:Sequence Analysis, DNA; D017381:Spliceosomes
chemical_list: D018841:HSP90 Heat-Shock Proteins; D009711:Nucleotides