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Molecular and genetic analysis of the sex-determining proteins FEM-1 and FEM-3.

Usha Vivegananthan

Molecular and genetic analysis of the sex-determining proteins FEM-1 and FEM-3.

by Usha Vivegananthan

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Written in English


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FEM-3, a protein with no recognizable functional motifs, acts as a developmental switch in sex determination. FEM-3 activity is both necessary and limiting for male development. Previous work showed that FEM-3 physically interacts with FEM-2, a type 2C protein phosphatase, but did not establish the biological significance of the interaction. I tested the hypothesis that the FEM-3/FEM-2 interaction is required for male development. Using a modified version of the yeast 2-hybrid system I isolated FEM-3(D313N), a mutant that is specifically defective in its ability to bind to FEM-2. Low-copy fem-3 transgenes that direct expression of the D313N variant under the control of native fem-3 regulatory sequences show significantly lower masculinizing activity than otherwise identical wild-type transgenes. These data suggest that the ability of FEM-3 to bind to FEM-2 is required for the masculinizing activity of FEM-3.Male development in Caenorhabditis elegans depends on the activities of the proteins FEM-1, FEM-2 and FEM-3. Extensive genetic analysis indicates that in XO animals the FEMs inhibit the activity of the female-determining transcription factor TRA-1A to promote male somatic development. In the germline, the FEMs act on TRA-1A and additional targets to promote spermatogenesis. Little is known about the molecular and biochemical mechanisms through which the FEMs act, especially the ankyrin (ANK) repeat protein FEM-1. I molecularly and genetically characterized a collection of fem-1 loss-of-function alleles and found that the N-terminal ANK repeats of FEM-1 are critical for its sex-determining activity. ANK repeats are evolutionarily conserved protein-protein interaction motifs suggesting that a FEM-1-ANK repeat mediated protein interaction is required for male development. Observations of a dominant-negative version of FEM-1 that carries a missense mutation in the sixth ANK repeat support this hypothesis. FEM-1 is unusual among the sex-determining proteins in that it has been well conserved through evolution: orthologues exist in insects and vertebrates. Others have suggested that the conservation of FEM-1 may be due to a role in apoptosis. However, my work demonstrates that FEM-1 is not essential for programmed cell death in C. elegans other than through its characterized role as a regulator of TRA-1A activity.

The Physical Object
Pagination208 leave.
Number of Pages208
ID Numbers
Open LibraryOL20338940M
ISBN 100612916324

cant advantage over isozyme analysis when large num-bers of markers are compared. Seed Storage Proteins—Seed storage proteins have also been used to compare genetic diversity (Fairbanks and others ). Seed storage proteins are deposited in relatively large quantities in mature seeds and typically remain stable until the seed germinates. Multiple Choice Questions on Molecular Genetics 1. The end product of translation are. a) amino acids a) DNA and protein coat. b) RNA and protein coat. c) Polysaccharides and proteins. d)a nucleic acid and a protein coat. 3. In bacteria, a small circle of DNA found outside the main chromosome is called a Autosomal Dominant vs X linked.

SdC- 1 fem- 1 sdc-3 fem-3 xx Off ON Off ON Off ON x0 ON Off ON Off ON Off (3 FIGURE The genetic regulatory pathway governing somatic sex determination in C. elegans. The genes fall into two classes: those that control sex determination and dosage compensation (xol-I, sdc-1, sdc-2 and sdc-3) and those that function exclusively. Genetic analysis suggests that tra-2 is a negative regulator of three genes that are required for male development: fem-1, fem-2, and fem We report that the carboxy-terminal region of TRA-2A.

Recent Molecular Genetics and Metabolism Articles Recently published articles from Molecular Genetics and Metabolism. Efficient engraftment of genetically modified cells is necessary to ameliorate central nervous system involvement of murine model of mucopolysaccharidosis type II by hematopoietic stem cell targeted gene therapy.   The product of fem-1, a nematode sex-determining gene, contains a motif found in cell cycle control proteins and receptors for cell-cell interactions. C Staden, R. ().


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Molecular and genetic analysis of the sex-determining proteins FEM-1 and FEM-3 by Usha Vivegananthan Download PDF EPUB FB2

Sex-determining protein; essential for the adoption of the male sexual fate in all tissues. Substrate recognition subunit of the cullin-RING-based CBC(fem-1) (Cul2-ElonginB-ElonginC) E3 ubiquitin-protein ligase complex which mediates in association with cofactors fem-2 and fem-3 the ubiquitination and subsequent proteasomal degradation of tra In Caenorhabditis elegans, the Gli-family transcription factor TRA-1 is the terminal effector of the sex-determination pathway.

TRA-1 activity inhibits male development and allows female fates. Genetic studies have indicated that TRA-1 is negatively regulated by the fem-1, fem-2, and fem-3 genes.

However, the mechanism of this regulation has not been by:   In the nematode Caenorhabditis elegans, fem-1, fem-2, and fem-3 play crucial roles in male sexual development. Among these three genes, fem-2 encodes a PP2C (serine/threonine phosphatase type 2C)-like protein, whose activity promotes the development of masculinity.

Different from the canonical PP2Cs, FEM-2 consists of an additional N-terminal domain (NTD) apart from its C Cited by: 7. Required for male development. In XO (male) animals, fem-3 directs male differentiation in all tissues.

In XX (hermaphrodite) animals, it specifies the first 80 or so germ cells to be sperm. Negatively regulates male development when bound to tra Together with fem-2 associates with the CBC(fem-1) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent.

Caenorhabditis elegans sex-determining protein FEM-2 is a protein phosphatase that promotes male development and interacts directly with FEM-3 Ian D. Chin-Sang and Andrew M. Spence 1 Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada, M5S 1A8. We report the cloning and sequencing of fem-1, a gene required for sex determination in both germline and somatic tissues in the nematode C.

elegans. Clones carrying a kb fragment are able to rescue the progeny of a fem-1 mutant when injected into its oocytes. The major fem-1 transcript in both sexes is kb and comprises 11 exons.

It encodes a soluble, intracellular protein of amino. FEM-3 —It was reported that three FEM proteins (FEM-1, -2, and -3) could interact with each other in vivo to constitute a functional FEM-1/2/3 complex, mediating male sexual devel. tra-2 (Doniach ), and fem-3 (Barton et al.

Genes with If and gf alleles that result in opposite trans­ formation phenotypes are likely to be central 'switch genes' in the pathway of sex determination.

Among the sex-determining genes of C. elegans, three fem genes, fem-1, fem-2, and fem The NTD Mediates the Binding of FEM-2 to FEM-1 and FEM It was reported that three FEM proteins (FEM-1, -2, and -3) could interact with each other in vivo to constitute a functional FEM-1/2/3 complex, mediating male sexual development.

Thus, we proposed that the NTD of FEM-2 might participate in protein-protein interaction in the assembly of. Abstract. The tra-2 gene of the nematode Caenorhabditis elegans encodes a predicted membrane protein, TRA-2A, that promotesXX hermaphrodite development.

Genetic analysis suggests thattra-2 is a negative regulator of three genes that are required for male development: fem-1, fem-2, and femWe report that the carboxy-terminal region of TRA-2A interacts specifically with FEM-3 in the yeast two.

stroyed either protein or RNA. However, treatments that destroyed DNA eliminated the transforming activity (Figure ). These experiments implied that the substance re-sponsible for genetic transformation was the DNA of the cell—hence that DNA is the genetic material. 4 Chapter 1 Introduction to Molecular Genetics and Genomics Living S cells.

Somatic sex determination in Caenorhabditis elegans involves a signal transduction pathway linking a membrane receptor to a transcription factor. The fem-2 gene is central to this pathway, producing a protein phosphatase (FEM-2) of the type 2C (PP2C). FEM-2 contains a long amino terminus that is absent in canonical PP2C enzymes.

The function of this domain is difficult to predict, since it. The product of fem-1, a nematode sex-determining gene, contains a motif found in cell cycle control proteins and receptors for cell-cell interactions.

Cell. Mar 23; 60 (6)– Stone JM, Collinge MA, Smith RD, Horn MA, Walker JC. Interaction of a protein phosphatase with an Arabidopsis serine-threonine receptor kinase. Science. The feminization-1c (fem-1c) gene has been shown to be associated with sex differentiation and determination in many metazoan species.

It belongs to the fem-1 family which is a member of the ANK superfamily. In this study, the full-length cDNA of the fem-1c (Hcfem-1c) gene was isolated from the freshwater mussel (Hypriopsis cumingii).The isolated Hcfem-1c cDNA was bp. Genetic studies have identified fem-1, fem-2, and fem-3 as immediate upstream negative regulators of tra-1 (Doniach and Hodgkin,Hodgkin,Kimble et al., ).

Loss of fem-1, fem-2, or fem-3 leads to increased TRA-1A levels, indicating that all three FEM proteins are required to negatively regulate TRA-1 levels in C. elegans. When. Genetic studies have identified fem-1, fem-2, and fem-3 as immediate upstream negative regulators of tra-1 (Doniach and Hodgkin, ; Hodgkin, ; Kimble et al., ).

Loss of fem-1, fem-2, or fem-3 leads to increased TRA-1A levels, indicating that all three FEM proteins are required to negatively regulate TRA-1 levels in C. elegans.

When. The feminizing suppressors are mostly alleles of known sex-determining genes: tra-1 (11 dominant alleles), tra-2 (one dominant allele), fem-1 (four alleles) and fem-2 (four alleles), but 18 are alleles of a new gene, fem Additional alleles have been isolated for the fem-2 and fem-3 genes, as well as fem-3 deficiencies.

The tra-2 gene of the nematode Caenorhabditis elegans encodes a predicted membrane protein, TRA-2A, that promotes XX hermaphrodite development. Genetic analysis suggests that tra-2 is a negative regulator of three genes that are required for male development: fem-1, fem-2, and femWe report that the carboxy-terminal region of TRA-2A interacts specifically with FEM-3 in the yeast two.

The tra-1 gene is the terminal control gene for somatic sex determination in the nematode Caenorhabditis elegans. Here we identify two tra-1 mRNAs: one is a kb transcript that peaks in abundance in the second larval stage, and the other is a 5 kb transcript that is present at relatively constant abundance throughout development.

Both RNAs occur at similar levels in both sexes, suggesting. Rosenquist TA, Kimble J. Molecular cloning and transcript analysis of fem-3, a sex-determination gene in Caenorhabditis elegans. Genes Dev. May; 2 (5)– Kuwabara PE. Interspecies comparison reveals evolution of control regions in the nematode sex-determining gene tra Genetics.

Oct; (2)– [PMC free article]. We have isolated nine gain-of-function (gf) alleles of the sex-determination gene fem-3 as suppressors of feminizing mutations in fem-1 and femThe wild-type fem-3 gene is needed for spermatogenesis in XX self-fertilizing hermaphrodites and for male development in both soma and germ line of XO animals.

Loss-of-function alleles of fem-3 transform XX and XO animals into females (spermless.In Caenorhabditis elegans,fem-1, fem-2, and fem-3 play pivotal roles in sex determination. Recently, a mammalian homologue of the C.

elegans sex-determining protein FEM-1, F1Aα, has been described. We report the cloning and sequencing of fem-1, a gene required for sex determination in both germline and somatic tissues in the nematode C. carrying a kb fragment are able to rescue the progeny of a fem-1 mutant when injected into its oocytes.

The major fem-1 transcript in both sexes is kb and comprises 11 exons. It encodes a soluble, intracellular protein of .