Identification of a secreted casein kinase 1 in Leishmania donovani: effect of protein over expression on parasite growth and virulence

Casein kinase 1 (CK1) plays an important role in eukaryotic signaling pathways, and their substrates include key regulatory proteins involved in cell differentiation, proliferation and chromosome segregation. The Leishmania genome encodes six potential CK1 isoforms, of which five have orthologs in o...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2013-11, Vol.8 (11), p.e79287-e79287
Main Authors: Dan-Goor, Mary, Nasereddin, Abedelmajeed, Jaber, Hanan, Jaffe, Charles L
Format: Article
Language:English
Subjects:
Analysis
Animals
Antibodies
Casein
Casein kinase I
Casein Kinase I - biosynthesis
Casein Kinase I - genetics
Cell culture
Cell differentiation
Cytometry
Cytosol
Differentiation (biology)
E coli
Flow cytometry
Gene Expression Regulation, Enzymologic
Genomes
Genomics
Growth
Health aspects
Immunofluorescence
Isoforms
Kinases
Leishmania
Leishmania donovani
Leishmania donovani - enzymology
Leishmania donovani - genetics
Leishmania donovani - pathogenicity
Leishmaniasis, Visceral - enzymology
Leishmaniasis, Visceral - genetics
Macrophages
Macrophages, Peritoneal - enzymology
Macrophages, Peritoneal - parasitology
Macrophages, Peritoneal - pathology
Mice
Mice, Inbred BALB C
Parasites
Parasitic diseases
Peritoneum
Promastigotes
Protein kinase
Protein kinases
Protein Structure, Tertiary
Proteins
Protozoan Proteins - biosynthesis
Protozoan Proteins - genetics
Regulatory proteins
Signaling
Substrates
Therapeutic applications
Tropical diseases
Viral antibodies
Virulence
Virulence (Microbiology)
Virulence Factors - biosynthesis
Virulence Factors - genetics
Western blotting
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Casein kinase 1 (CK1) plays an important role in eukaryotic signaling pathways, and their substrates include key regulatory proteins involved in cell differentiation, proliferation and chromosome segregation. The Leishmania genome encodes six potential CK1 isoforms, of which five have orthologs in other trypanosomatidae. Leishmania donovani CK1 isoform 4 (Ldck1.4, orthologous to LmjF27.1780) is unique to Leishmania and contains a putative secretion signal peptide. The full-length gene and three shorter constructs were cloned and expressed in E. coli as His-tag proteins. Only the full-length 62.3 kDa protein showed protein kinase activity indicating that the N-terminal and C-terminal domains are essential for protein activity. LdCK1.4-FLAG was stably over expressed in L. donovani, and shown by immunofluorescence to be localized primarily in the cytosol. Western blotting using anti-FLAG and anti-CK1.4 antibodies showed that this CK1 isoform is expressed and secreted by promastigotes. Over expression of LdCK1.4 had a significant effect on promastigote growth in culture with these parasites growing to higher cell densities than the control parasites (wild-type or Ld:luciferase, P
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0079287