CP beta3, a novel isoform of an actin-binding protein, is a component of the cytoskeletal calyx of the mammalian sperm head.
In the mammalian sperm head, the nucleus is tightly associated with the calyx, a cell type-specific cytoskeletal structure. Previously, we have identified and characterized some basic proteins such as calicin and cylicins I and II as major calyx components of bovine and human spermatids and spermatozoa. Surprisingly we have now discovered another calyx constituent which by amino acid sequencing and cDNA cloning was recognized as a novel isoform of the widespread beta subunit of the heterodimeric actin-binding "capping protein" (CP). This polypeptide, CP beta3, of sperm calices, is identical with the beta2 subunit present in diverse somatic cell types, except that it shows an amino-terminal extension of 29 amino acids and its mRNA is detected only in testis and, albeit in trace amounts, brain. This CP beta3 mRNA contains the additional sequence, encoded by exon 1 of the gene, which is missing in beta2 mRNAs. Antibodies specific for the beta3 amino-terminal addition have been used to identify the protein by immunoblotting and to localize it to the calyx structure by immunofluorescence microscopy. We conclude that in spermiogenesis the transcription of the gene encoding the beta1, beta2, and beta3 CP subunits is regulated specifically to include exon 1 and to give rise to the testis isoform CP beta3, which is integrated into the calyx structure of the forming sperm head. This surprising finding of an actin-binding protein isoform in an insoluble cytoskeletal structure is discussed in relation to the demonstrated roles of actin and certain actin-binding proteins, such as Limulus alpha-scruin, in spermiogenesis and spermatozoa.
von Bülow
M
M
Division of Cell Biology, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
Rackwitz
H R
HR
Zimbelmann
R
R
Franke
W W
WW
D051339:Actin Depolymerizing Factors; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry / Q000235:genetics; D002417:Cattle; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000737:chemistry*; D051345:Destrin; D005455:Fluorescent Antibody Technique; D008297:Male; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D017422:Sequence Analysis, DNA; D013077:Sperm Head / Q000737:chemistry*; D013094:Spermatozoa / Q000166:cytology
D051339:Actin Depolymerizing Factors; D051345:Destrin; D008840:Microfilament Proteins; D012333:RNA, Messenger