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PMID:27624483

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Citation

Young, SA, Miyata, H, Satouh, Y, Muto, M, Larsen, MR, Aitken, RJ, Baker, MA and Ikawa, M (2016) CRISPR/Cas9-mediated mutation revealed cytoplasmic tail is dispensable for IZUMO1 function and male fertility. Reproduction 152:665-672

Abstract

IZUMO1 is a protein found in the head of spermatozoa that has been identified as essential for sperm-egg fusion. Its binding partner in the egg has been discovered (JUNO); however, the roles of several domains within IZUMO1 remain unexplored. One such domain is the C-terminus, which undergoes major phosphorylation changes in the cytoplasmic portion of the protein during rat epididymal transit. However, the cytoplasmic tail of IZUMO1 in many species is highly variable, ranging from 55 to one amino acid. Therefore, to understand the role of the cytoplasmic tail of IZUMO1 in mouse, we utilised the gene manipulation system of CRISPR/Cas9 to generate a point mutation resulting in a premature stop codon, producing mice with truncated IZUMO1. Mice without the cytoplasmic tail of IZUMO1 showed normal fertility but decreased the amount of protein, indicating that whilst this region is important for the expression level of IZUMO1, it is dispensable for fertilisation in the mouse.

Links

PubMed Online version:10.1530/REP-16-0150

Keywords

Amino Acid Sequence; Animals; CRISPR-Cas Systems; Cytoplasm/metabolism; Fertility/genetics; Fertilization/physiology; Immunoglobulins/genetics; Immunoglobulins/metabolism; Male; Membrane Proteins/genetics; Membrane Proteins/metabolism; Mice; Mutation; Phosphorylation; Protein Domains; Sperm-Ovum Interactions; Spermatozoa/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:IZUM1

located_in

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IZUM1

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IZUM1

part_of

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IZUM1

part_of

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

Notes

See also

References

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