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. 2008 Oct 15;322(2):322-31.
doi: 10.1016/j.ydbio.2008.07.035. Epub 2008 Aug 7.

SKR-1, a homolog of Skp1 and a member of the SCF(SEL-10) complex, regulates sex-determination and LIN-12/Notch signaling in C. elegans

Darrell J Killian  1 , Elizabeth Harvey, Peter Johnson, Muneyoshi Otori, Shohei Mitani, Ding Xue
Affiliations

"VSports app下载" SKR-1, a homolog of Skp1 and a member of the SCF(SEL-10) complex, regulates sex-determination and LIN-12/Notch signaling in C. elegans

Darrell J Killian et al. Dev Biol. .

Abstract

Sex-determination in Caenorhabditis elegans requires regulation of gene transcription and protein activity and stability. sel-10 encodes a WD40-repeat-containing F-box protein that likely mediates the ubiquitin-mediated degradation of important sex-determination factors. Loss of sel-10 results in a mild masculinization of hermaphrodites, whereas dominant alleles of sel-10, such as sel-10(n1074), cause a more severe masculinization, including a reversal of the life versus death decision in sex-specific neurons. To investigate about how sel-10 regulates sex-determination, we conducted a sel-10(n1074) suppressor screen and isolated a weak loss-of-function allele of skr-1, one of 21 Skp1-related genes in C. elegans. Skp1, Cullin, and F-box proteins, such as SEL-10, are components of the SCF E3 ubiquitin-ligase complex. We present genetic evidence that the sel-10(n1074) masculinization phenotype is dependent upon skr-1 and cul-1 activity. Furthermore, we show that the SKR-1(M140I) weak loss-of-function mutation interferes with SKR-1/SEL-10 binding. Unexpectedly, we found that the G567E substitution in SEL-10 caused by the n1074 allele impairs the binding of SEL-10 to SKR-1 and the dimerization of SEL-10, which may be important for SEL-10 function. Our results suggest that SKR-1, CUL-1 and SEL-10 constitute an SCF E3 ligase complex that plays an important role in modulating sex-determination and LIN-12/Notch signaling in C VSports手机版. elegans. .

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"VSports手机版" Figures

Figure 1
Figure 1
A simplified view of the sex-determination pathway in XX animals (A) and XO animals (B). Sex-determination genes depicted in black are active and genes depicted in grey are repressed (see text for detail). SCFSEL-10 is an E3 ubiquitin ligase composed of SKR-1, CUL-1, and SEL-10.
Figure 2
Figure 2
(A) Schematic representation of the skr-2 and skr-1 locus of chromosome I (LG: I). The skr-1(sm151) lesion is indicated by an arrowhead and the skr-1(tm2391) deletion is indicated by a dashed line. Green lines represent PCR fragments that rescued skr-1(sm151) and the red lines represent PCR fragments that did not rescue. (B) Alignment was performed between Skp1-related proteins in human (Hs), mouse (Mm), zebrafish (Dr), fruitfly (Dm), worm (Ce), fission yeast (Sp), and budding yeast (Sc) with ClustalW2 (European Bioinformatics Institute of the European Molecular Biology Laboratory). (*) identical residues, (:) conserved residues, (.) semi-conserved residues. C. elegans SKR-1 M140 is indicated in red. F-box binding residues of human Skp1 are indicated in blue based on Schulmann et al. (2000).
Figure 3
Figure 3
DIC lateral views of (A) N2 normal uterine development and (B) skr-1(tm2391) uterine hyperplasia (ut = uterus). DIC ventral views of (C) N2 normal somatic gonad development and (D) skr- 1(tm2391) somatic gonad hyperplasia (sg = somatic gonad). Bars = 25 µm.
Figure 4
Figure 4
(A) A yeast two-hybrid assay with AD-SKR-1, DB-SEL-10, and the respective M140I and G567E mutants or empty vectors (−). Yeast were plated in a 0.5x dilution series from left to right starting at OD600 = 10. Positive interactions exhibit increased growth on 3AT and decreased growth on 5FOA relative to negative controls (lanes 5–8). Positive interactions are shown by lacZ staining. ®-galactosidase activity based on the CPRG assay is shown as a percentage of the activity observed in yeast expressing SKR-1(+)/SEL-10(+). Yeast were grown for two days before imaging. (B) Western blot analysis of yeast lysates containing AD-SKR-1 and AD-SKR-1(M140I) (left) and AD-SEL-10 and AD-SEL-10(G567E) (right). Phosphoglycerate kinase (PGK) was used as a loading control. See Materials and Methods for additional details.
Figure 5
Figure 5
A yeast two-hybrid assay with AD-SKR-1 or AD-SKR-1(M140) and DB-CUL-1 or empty vectors (−).Yeast were plated in a 0.5x dilution series from left to right starting at OD600 = 10. Positive interactions exhibit increased growth on 3AT relative to negative controls (lanes 1, 2, and 5). ®-galactosidase activity based on CPRG assay is shown as a percentage of the activity observed in yeast expressing SKR-1(+)/CUL-1(+). Yeast were grown for two days before imaging. See Materials and Methods for additional details.
Figure 6
Figure 6
A yeast two-hybrid assay with AD-SEL-10 and DB-SEL-10. Yeast were plated in a 0.5x dilution series from left to right starting at OD600 = 10. Positive interactions exhibit increased growth on 3AT relative to negative controls (lanes 5–8). Yeast were grown for seven days before imaging. See Materials and Methods for additional details.
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References (V体育2025版)

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