Cloning, characterization and sub-cellular localization of gamma subunit of T-complex protein-1 (chaperonin) from Leishmania donovani

• Published in: Biochemical and biophysical research communications 2012-12, Vol.429 (1-2), p.70-74
• Main Authors: Bhaskar, Kumari, Neeti, Goyal, Neena
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; ACTIN; Actins - metabolism; Amino acid sequence; Branches; Chaperonin; Chaperonin Containing TCP-1 - classification; Chaperonin Containing TCP-1 - genetics; Chaperonin Containing TCP-1 - metabolism; Chaperonins; CLONING; Cloning, Molecular; CYTOPLASM; Cytoskeleton; Differential expression; Flagella; GENES; Immunofluorescence; Leishmania donovani; Leishmania donovani - genetics; Leishmania donovani - metabolism; Log phase; MICROSCOPY; MICROTUBULES; Nuclei; PARASITES; Phylogeny; Promastigotes; Protein folding; Protein Structure, Tertiary; Protozoan Proteins - classification; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Stationary phase; STRUCTURAL CHEMICAL ANALYSIS; T-complex protein-1
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2012.10.090

► The study presents cloning & characterization of TCP1γ gene from L. donovani. ► TCP1γ is a subunit of T-complex protein-1 (TCP1), a chaperonin class of protein. ► LdTCPγ exhibited differential expression in different stages of promastigotes. ► LdTCPγ co-localized with actin, a cytoskeleton protein. ► The data suggests that this gene may have a role in differentiation/biogenesis. ► First report on this chapronin in Leishmania. T-complex protein-1 (TCP1) complex, a chaperonin class of protein, ubiquitous in all genera of life, is involved in intracellular assembly and folding of various proteins. The gamma subunit of TCP1 complex (TCP1γ), plays a pivotal role in the folding and assembly of cytoskeleton protein(s) as an individual or complexed with other subunits. Here, we report for the first time cloning, characterization and expression of the TCP1γ of Leishmania donovani (LdTCP1γ), the causative agent of Indian Kala-azar. Primary sequence analysis of LdTCP1γ revealed the presence of all the characteristic features of TCP1γ. However, leishmanial TCP1γ represents a distinct kinetoplastid group, clustered in a separate branch of the phylogenic tree. LdTCP1γ exhibited differential expression in different stages of promastigotes. The non-dividing stationary phase promastigotes exhibited 2.5-fold less expression of LdTCP1γ as compared to rapidly dividing log phase parasites. The sub-cellular distribution of LdTCP1γ was studied in log phase promastigotes by employing indirect immunofluorescence microscopy. The protein was present not only in cytoplasm but it was also localized in nucleus, peri-nuclear region, flagella, flagellar pocket and apical region. Co-localization of LdTCP1γ with actin suggests that, this gene may have a role in maintaining the structural dynamics of cytoskeleton of parasite.