Heme uptake by Leishmania amazonensis is mediated by the transmembrane protein LHR1.
Trypanosomatid protozoan parasites lack a functional heme biosynthetic pathway, so must acquire heme from the environment to survive. However, the molecular pathway responsible for heme acquisition by these organisms is unknown. Here we show that L. amazonensis LHR1, a homolog of the C. elegans plasma membrane heme transporter HRG-4, functions in heme transport. Tagged LHR1 localized to the plasma membrane and to endocytic compartments, in both L. amazonensis and mammalian cells. Heme deprivation in L. amazonensis increased LHR1 transcript levels, promoted uptake of the fluorescent heme analog ZnMP, and increased the total intracellular heme content of promastigotes. Conversely, deletion of one LHR1 allele reduced ZnMP uptake and the intracellular heme pool by approximately 50%, indicating that LHR1 is a major heme importer in L. amazonensis. Viable parasites with correct replacement of both LHR1 alleles could not be obtained despite extensive attempts, suggesting that this gene is essential for the survival of promastigotes. Notably, LHR1 expression allowed Saccharomyces cerevisiae to import heme from the environment, and rescued growth of a strain deficient in heme biosynthesis. Syntenic genes with high sequence identity to LHR1 are present in the genomes of several species of Leishmania and also Trypanosoma cruzi and Trypanosoma brucei, indicating that therapeutic agents targeting this transporter could be effective against a broad group of trypanosomatid parasites that cause serious human disease.
Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland, United States of America.
Chau
C
Huynh
Xiaojing
X
Yuan
Danilo C
DC
Miguel
Rebecca L
RL
Renberg
Olga
O
Protchenko
Caroline C
CC
Philpott
Iqbal
I
Hamza
Norma W
NW
Andrews
D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D006367:HeLa Cells; D006418:Heme / Q000172:deficiency / Q000378:metabolism*; D006801:Humans; D007894:Leishmania mexicana / Q000378:metabolism* / Q000472:pathogenicity; D008264:Macrophages / Q000378:metabolism / Q000469:parasitology; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008665:Metalloporphyrins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
D026901:Membrane Transport Proteins; D008665:Metalloporphyrins; D015800:Protozoan Proteins; D006418:Heme; C058069:zinc mesoporphyrin