Conservation in the 5′ flanking sequences of transcribed members of the Caenorhabditis elegans major sperm protein gene family

Caenorhabditis elegans has 12 tRNA(UGGTrp) genes as defined by Southern analysis. In order to evaluate the function of the individual members of this multigene family, we sought to recover amber (UAG)-suppressing mutations from reversion experiments with animals carrying amber mutations in a nervous system-affecting gene (unc-13) or a sex-determining gene (tra-3). Revertants were analyzed by Southern blot, exploiting the fact that the CCA to CTA change at the anticodon creates a new XbaI site. Five different members of the tRNATrp gene family were identified as suppressors: sup-7 X, sup-5 III, sup-24 IV, sup-28 X, and sup-29 IV. All five suppressor genes were sequenced and found to encode identical tRNA(UAGTrp) molecules with a single base change (CCA to CTA) at the anticodon compared with their wild-type counterparts. The flanking sequences had only limited homology. The relative expression of these five genes was determined by measuring the efficiencies of suppressers against amber mutations in genes affecting the nervous system, hypodermis, muscle, and sex determination. The results of these cross-suppression tests showed that the five members of the tRNA(Trp) gene family were differentially regulated in a tissue- or development stage-specific manner.

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2.6Å Resolution Crystal Structure of Helices of the Motile Major Sperm Protein (MSP) of Caenorhabditis elegans

Journal of Molecular Biology ◽

10.1016/s0022-2836(02)00294-2 ◽

2002 ◽

Vol 319 (2) ◽

pp. 491-499 ◽

Cited By ~ 21

Author(s):

Anne M.E. Baker ◽

Thomas M. Roberts ◽

Murray Stewart

Keyword(s):

Crystal Structure ◽

Caenorhabditis Elegans ◽

Major Sperm Protein ◽

Sperm Protein

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

Developmental Biology ◽

10.1016/0012-1606(81)90398-5 ◽

1981 ◽

Vol 84 (2) ◽

pp. 299-312 ◽

Cited By ~ 68

Author(s):

Michael R. Klass ◽

David Hirsh

Keyword(s):

Caenorhabditis Elegans ◽

Biochemical Analysis ◽

Major Sperm Protein ◽

Sperm Protein

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Isolation and characterization of a sperm-specific gene family in the nematode Caenorhabditis elegans

Molecular and Cellular Biology ◽

10.1128/mcb.4.3.529-537.1984 ◽

1984 ◽

Vol 4 (3) ◽

pp. 529-537

Author(s):

M R Klass ◽

S Kinsley ◽

L C Lopez

Keyword(s):

Caenorhabditis Elegans ◽

Gene Family ◽

Specific Gene ◽

Cdna Clones ◽

Major Sperm Protein ◽

Adult Males ◽

Flanking Sequence ◽

Nematode Caenorhabditis Elegans ◽

Developmentally Regulated ◽

Isolation And Characterization

The major sperm protein (MSP) of the nematode Caenorhabditis elegans is a low-molecular-weight (15,000) basic protein implicated in the pseudopodial movement of mature spermatozoa. Its synthesis occurs in a specific region of the gonad and is regulated at the level of transcription (M. Klass and D. Hirsh, Dev. Biol. 84:299-312, 1981; S. Ward and M. Klass, Dev. Biol. 92:203-208, 1982; Klass et al., Dev. Biol. 93:152-164, 1982). A developmentally regulated gene family has been identified that codes for this MSP. Whole genomic blots, as well as analysis of genomic clone banks, indicate that there are between 15 and 25 copies of the MSP gene in the nematode genome. Southern blot analysis also indicates that there is no rearrangement or amplification within the MSP gene family during development. No evidence was found of methylation at various restriction sites surrounding the MSP gene family, and similarly, no correlation between methylation and expression was observed. Three distinct members of this MSP gene family have been cloned, and their nucleotide sequences have been determined. Differential screening of a cDNA clone bank made from polyadenylated mRNA from adult males yielded 45 male-specific clones, 32 of which were clones of MSP genes. One of these cDNA clones was found to contain the entire nucleotide sequence for the MSP, including part of the 5' leader and all of the 3' trailing sequence. Genomic clones bearing copies of the MSP genes have been isolated. At least one of the members of this gene family is a pseudogene. Another member of the MSP gene family that has been cloned from genomic DNA contains the entire uninterrupted structural sequence for the MSP in addition to a 5' flanking sequence containing a promoter-like region with the classic TATA box, a sequence resembling the CAAT box, and a putative ribosome-binding sequence. The 3' trailing sequences of the genomic and the cDNA clones contain an AATAAA polyadenylation site.

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Differential expression of five tRNA(UAGTrp) amber suppressors in Caenorhabditis elegans

Molecular and Cellular Biology ◽

10.1128/mcb.8.9.3627-3635.1988 ◽

1988 ◽

Vol 8 (9) ◽

pp. 3627-3635 ◽

Cited By ~ 1

Author(s):

K Kondo ◽

J Hodgkin ◽

R H Waterston

Keyword(s):

Nervous System ◽

Caenorhabditis Elegans ◽

Gene Family ◽

Development Stage ◽

Base Change ◽

Wild Type ◽

Specific Manner ◽

Flanking Sequences ◽

Single Base Change ◽

The Individual

Caenorhabditis elegans has 12 tRNA(UGGTrp) genes as defined by Southern analysis. In order to evaluate the function of the individual members of this multigene family, we sought to recover amber (UAG)-suppressing mutations from reversion experiments with animals carrying amber mutations in a nervous system-affecting gene (unc-13) or a sex-determining gene (tra-3). Revertants were analyzed by Southern blot, exploiting the fact that the CCA to CTA change at the anticodon creates a new XbaI site. Five different members of the tRNATrp gene family were identified as suppressors: sup-7 X, sup-5 III, sup-24 IV, sup-28 X, and sup-29 IV. All five suppressor genes were sequenced and found to encode identical tRNA(UAGTrp) molecules with a single base change (CCA to CTA) at the anticodon compared with their wild-type counterparts. The flanking sequences had only limited homology. The relative expression of these five genes was determined by measuring the efficiencies of suppressers against amber mutations in genes affecting the nervous system, hypodermis, muscle, and sex determination. The results of these cross-suppression tests showed that the five members of the tRNA(Trp) gene family were differentially regulated in a tissue- or development stage-specific manner.

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

Developmental Biology ◽

10.1016/j.ydbio.2006.07.013 ◽

2006 ◽

Vol 299 (1) ◽

pp. 105-121 ◽

Cited By ~ 43

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

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Isolation and characterization of a sperm-specific gene family in the nematode Caenorhabditis elegans.

Molecular and Cellular Biology ◽

10.1128/mcb.4.3.529 ◽

1984 ◽

Vol 4 (3) ◽

pp. 529-537 ◽

Cited By ~ 31

Author(s):

M R Klass ◽

S Kinsley ◽

L C Lopez

Keyword(s):

Caenorhabditis Elegans ◽

Gene Family ◽

Specific Gene ◽

Cdna Clones ◽

Major Sperm Protein ◽

Adult Males ◽

Flanking Sequence ◽

Nematode Caenorhabditis Elegans ◽

Developmentally Regulated ◽

Isolation And Characterization

The major sperm protein (MSP) of the nematode Caenorhabditis elegans is a low-molecular-weight (15,000) basic protein implicated in the pseudopodial movement of mature spermatozoa. Its synthesis occurs in a specific region of the gonad and is regulated at the level of transcription (M. Klass and D. Hirsh, Dev. Biol. 84:299-312, 1981; S. Ward and M. Klass, Dev. Biol. 92:203-208, 1982; Klass et al., Dev. Biol. 93:152-164, 1982). A developmentally regulated gene family has been identified that codes for this MSP. Whole genomic blots, as well as analysis of genomic clone banks, indicate that there are between 15 and 25 copies of the MSP gene in the nematode genome. Southern blot analysis also indicates that there is no rearrangement or amplification within the MSP gene family during development. No evidence was found of methylation at various restriction sites surrounding the MSP gene family, and similarly, no correlation between methylation and expression was observed. Three distinct members of this MSP gene family have been cloned, and their nucleotide sequences have been determined. Differential screening of a cDNA clone bank made from polyadenylated mRNA from adult males yielded 45 male-specific clones, 32 of which were clones of MSP genes. One of these cDNA clones was found to contain the entire nucleotide sequence for the MSP, including part of the 5' leader and all of the 3' trailing sequence. Genomic clones bearing copies of the MSP genes have been isolated. At least one of the members of this gene family is a pseudogene. Another member of the MSP gene family that has been cloned from genomic DNA contains the entire uninterrupted structural sequence for the MSP in addition to a 5' flanking sequence containing a promoter-like region with the classic TATA box, a sequence resembling the CAAT box, and a putative ribosome-binding sequence. The 3' trailing sequences of the genomic and the cDNA clones contain an AATAAA polyadenylation site.

Download Full-text