Sperm isolation and biochemical analysis of the major sperm protein from Caenorhabditis elegans

1981 ◽  
Vol 84 (2) ◽  
pp. 299-312 ◽  
Author(s):  
Michael R. Klass ◽  
David Hirsh
Keyword(s):  
Caenorhabditis Elegans ◽  
Biochemical Analysis ◽  
Major Sperm Protein ◽  
Sperm Protein

scholarly journals Major sperm protein signaling promotes oocyte microtubule reorganization prior to fertilization in Caenorhabditis elegans

2006 ◽  
Vol 299 (1) ◽  
pp. 105-121 ◽  
Author(s):  
Jana E. Harris ◽  
J. Amaranath Govindan ◽  
Ikuko Yamamoto ◽  
Joel Schwartz ◽  
Irina Kaverina ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Protein Signaling ◽  
Major Sperm Protein ◽  
Sperm Protein

scholarly journals The Caenorhabditis elegans spe-6 gene is required for major sperm protein assembly and shows second site non-complementation with an unlinked deficiency.

Genetics ◽  
1993 ◽  
Vol 133 (1) ◽  
pp. 79-86 ◽  
Author(s):  
J P Varkey ◽  
P L Jansma ◽  
A N Minniti ◽  
S Ward
Keyword(s):  
Caenorhabditis Elegans ◽  
Gene Product ◽  
Spindle Pole ◽  
Specific Gene ◽  
Major Sperm Protein ◽  
Sperm Protein ◽  
Fibrous Body ◽  
Sperm Development ◽  
Chromosome Iii

Abstract Caenorhabditis elegans spermatozoa move by crawling. Their motility requires thin cytoskeletal filaments assembled from a unique cytoskeletal protein, the major sperm protein (MSP). During normal sperm development the MSP is segregated to developing sperm by assembly into filaments that form a paracrystalline array in a transient organelle, the fibrous body-membranous organelle. Mutations in the spe-6 gene cause sterility because they lead to defective primary spermatocytes that do not form spermatids. In these mutant spermatocytes the MSP fails to assemble into fibrous body filaments. Instead, the unassembled MSP distributes throughout the cytoplasm and nucleus. Thus, the spe-6 gene product is necessary for normal MSP localization and assembly during sperm development. In addition to their MSP assembly defect, spe-6 mutant spermatocytes arrest meiosis at diakinesis although their spindle pole bodies still replicate and separate. This results in spermatocytes with four half-spindles surrounding condensed, but unsegregated, chromosomes. All four spe-6 alleles, as well as a chromosome III deficiency that deletes the spe-6 gene, fail to complement two small overlapping chromosome IV deficiencies, eDf18 and eDf19. This non-allele-specific second site non-complementation suggests a concentration-dependent interaction between the spe-6 gene product and products of the gene(s) under eDf18 and eDf19, which include a cluster of sperm-specific genes. Since MSP filament assembly is highly concentration-dependent in vitro, the non-complementation might be expected if the sperm-specific gene products under eDf18 and eDf19 were needed together with the spe-6 gene product to promote MSP assembly.

Genomic organization of major sperm protein genes and pseudogenes in the nematode Caenorhabditis elegans

1988 ◽  
Vol 199 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Samuel Ward ◽  
Daniel J. Burke ◽  
John E. Sulston ◽  
Alan R. Coulson ◽  
Donna G. Albertson ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Genomic Organization ◽  
Major Sperm Protein ◽  
Nematode Caenorhabditis Elegans ◽  
Sperm Protein

Structure and macromolecular assembly of two isoforms of the major sperm protein (MSP) from the amoeboid sperm of the nematode, Ascaris suum

1992 ◽  
Vol 101 (4) ◽  
pp. 847-857 ◽  
Author(s):  
K.L. King ◽  
M. Stewart ◽  
T.M. Roberts ◽  
M. Seavy
Keyword(s):  
Polyethylene Glycol ◽  
Biochemical Analysis ◽  
Critical Concentration ◽  
Cytoskeletal Proteins ◽  
Amoeboid Movement ◽  
Major Sperm Protein ◽  
Sperm Protein ◽  
Macromolecular Assembly ◽  
Amoeboid Motility ◽  
Simple Alternative

Ascaris sperm are amoeboid cells that crawl by extending pseudopods. Although amoeboid motility is generally mediated through an actin-based cytoskeleton, Ascaris sperm lack this system. Instead, their major sperm protein (MSP) forms an extensive filament system that appears to fulfil this function. Because their motility appears to be essentially the same as that of their actin-rich counterparts, Ascaris sperm offer a simple alternative system for investigation of the molecular mechanism of amoeboid movement. To examine the structure and composition of the cytoskeleton, we stabilized the extremely labile native MSP filaments by detergent lysis of sperm in the presence of either glutaraldehyde or polyethylene glycol (PEG). Biochemical analysis showed that the cytoskeleton contained two isoforms of MSP, designated alpha- and beta-, that we purified and sequenced. Both contain 126 amino acids and have an acetylated N-terminal alanine, but differ at four residues so that alpha-MSP is 142 Da larger and 0.6 pH unit more basic than beta-MSP. Neither isoform shares sequence homology with other cytoskeletal proteins. In ethanol, 2-methyl-2,4-pentanediol (MPD), and other water-miscible alcohols each isoform assembled into filaments 10 nm wide with a characteristic substructure repeating axially at 9 nm. These filaments were indistinguishable from native fibers isolated from detergent-lysed sperm. Pelleting assays indicated a critical concentration for assembly of 0.2 mM for both isoforms in 30% ethanol, but alpha-MSP formed filaments at lower solvent concentration than beta-MSP. When incubated in polyethylene glycol, both isoforms formed thin, needle-shaped crystals that appeared to be constructed from helical fibers, with a 9 nm axial repeat that matched that seen in isolated filaments. These crystals probably contained a parallel array of helical filaments, and may enable both the structure of MSP molecules and their mode of assembly into higher aggregates to be investigated to high resolution.

scholarly journals Localized Depolymerization of the Major Sperm Protein Cytoskeleton Correlates with the Forward Movement of the Cell Body in the Amoeboid Movement of Nematode Sperm

1999 ◽  
Vol 146 (5) ◽  
pp. 1087-1096 ◽  
Author(s):  
Joseph E. Italiano ◽  
Murray Stewart ◽  
Thomas M. Roberts
Keyword(s):  
Cell Body ◽  
Body Movement ◽  
Leading Edge ◽  
Amoeboid Movement ◽  
Major Sperm Protein ◽  
Membrane Protrusion ◽  
Forward Movement ◽  
Sperm Protein ◽  
Amoeboid Motility ◽  
Tightly Coupled

The major sperm protein (MSP)-based amoeboid motility of Ascaris suum sperm requires coordinated lamellipodial protrusion and cell body retraction. In these cells, protrusion and retraction are tightly coupled to the assembly and disassembly of the cytoskeleton at opposite ends of the lamellipodium. Although polymerization along the leading edge appears to drive protrusion, the behavior of sperm tethered to the substrate showed that an additional force is required to pull the cell body forward. To examine the mechanism of cell body movement, we used pH to uncouple cytoskeletal polymerization and depolymerization. In sperm treated with pH 6.75 buffer, protrusion of the leading edge slowed dramatically while both cytoskeletal disassembly at the base of the lamellipodium and cell body retraction continued. At pH 6.35, the cytoskeleton pulled away from the leading edge and receded through the lamellipodium as its disassembly at the cell body continued. The cytoskeleton disassembled rapidly and completely in cells treated at pH 5.5, but reformed when the cells were washed with physiological buffer. Cytoskeletal reassembly occurred at the lamellipodial margin and caused membrane protrusion, but the cell body did not move until the cytoskeleton was rebuilt and depolymerization resumed. These results indicate that cell body retraction is mediated by tension in the cytoskeleton, correlated with MSP depolymerization at the base of the lamellipodium.

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