Examination of a novel head-stalk protein family in Giardia lamblia characterised by the pairing of ankyrin repeats and coiled-coil domains.
The intestinal pathogen Giardia lamblia possesses several unusual organelle features, including two equivalent nuclei, no mitochondria or peroxisomes, and a developmentally regulated rough endoplasmic reticulum and Golgi. Giardia also possesses a number of complex and unique cytoskeleton structures that dictate cell shape, motility and attachment. Our investigations of cytoskeletal proteins have revealed the presence of a new protein family. Proteins in this family contain both ankyrin repeats and coiled-coil domains; although these are common protein motifs, their pairing is unique, thus establishing a new class of head-stalk proteins. Examination of the G. lamblia genome shows evidence for at least 18 genes coding for proteins with a series of ankyrin repeats followed by a lengthy coiled-coil domain and at least an additional 14 genes coding for proteins with a prominent coiled-coil domain flanked by two series of ankyrin repeats. We have examined one of these proteins, Giardia Axoneme Associated Protein (GASP-180), in detail. GASP-180 is a 180 kDa protein containing five ankyrin repeats in a 200 amino acid N-terminal domain separated by a short spacer from an approximately 1375 amino acid coiled-coil domain. Using anti-peptide antibodies raised against a unique 20 amino acid sequence found at the C-terminus, we have determined that GASP-180 is present in cytoskeleton extractions of the parasite and localises to the proximal base of the anterior flagellar axonemes. The combination of the localisation and the structural and functional motifs of GASP-180 make it a strong candidate to participate in control of flagellar activity.
Biology Department, 348 Reiss Building, 37th and O Sts NW, Georgetown University, Washington, DC 20057, USA. hge@georgetown.edu
Heidi G
HG
Elmendorf
Sally C
SC
Rohrer
Rasha S
RS
Khoury
Rachel E
RE
Bouttenot
Theodore E
TE
Nash
D000595:Amino Acid Sequence; D000818:Animals; D017089:Ankyrin Repeat; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D003599:Cytoskeleton / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D005407:Flagella / Q000737:chemistry; D016829:Giardia lamblia / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
C058979:33 kDa structural protein, Giardia; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D016054:DNA, Protozoan; D015800:Protozoan Proteins