Identification of the centromeres of Leishmania major: revealing the hidden pieces

EMBO Rep. 2017 Nov;18(11):1968-1977. doi: 10.15252/embr.201744216. Epub 2017 Sep 21.

Abstract

Leishmania affects millions of people worldwide. Its genome undergoes constitutive mosaic aneuploidy, a type of genomic plasticity that may serve as an adaptive strategy to survive distinct host environments. We previously found high rates of asymmetric chromosome allotments during mitosis that lead to the generation of such ploidy. However, the underlying molecular events remain elusive. Centromeres and kinetochores most likely play a key role in this process, yet their identification has failed using classical methods. Our analysis of the unconventional kinetochore complex recently discovered in Trypanosoma brucei (KKTs) leads to the identification of a Leishmania KKT gene candidate (LmKKT1). The GFP-tagged LmKKT1 displays "kinetochore-like" dynamics of intranuclear localization throughout the cell cycle. By ChIP-Seq assay, one major peak per chromosome is revealed, covering a region of 4 ±2 kb. We find two largely conserved motifs mapping to 14 of 36 chromosomes while a higher density of retroposons are observed in 27 of 36 centromeres. The identification of centromeres and of a kinetochore component of Leishmania chromosomes opens avenues to explore their role in mosaic aneuploidy.

Keywords: Leishmania; ChIP‐sequencing; centromeres; fluorescent in situ hybridization; kinetoplastid kinetochores.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aneuploidy
  • Base Sequence
  • Centromere / metabolism*
  • Centromere / ultrastructure
  • Chromatin Immunoprecipitation
  • Chromosome Mapping
  • Chromosomes / chemistry*
  • Gene Expression
  • Genome, Protozoan*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • High-Throughput Nucleotide Sequencing
  • Kinetochores / metabolism*
  • Kinetochores / ultrastructure
  • Leishmania major / genetics*
  • Leishmania major / metabolism
  • Mitosis
  • Mosaicism
  • Protozoan Proteins / genetics*
  • Protozoan Proteins / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Protozoan Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins