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Sucrose phosphate synthase and sucrose phosphate phosphatase interact in planta and promote plant growth and biomass accumulation.

Maloney VJ, Park JY, Unda F, Mansfield SD - J. Exp. Bot. (2015)

Bottom Line: Herein, the formation of an enzyme complex between SPS and SPP was examined, and the results from yeast two-hybrid experiments suggest that there is indeed an association between these proteins.The findings clearly demonstrated that SPS interacts with SPP and that this interaction impacts soluble carbohydrate pools and affects carbon partitioning to starch.Moreover, a fusion construct between the two genes promotes plant growth in both transgenic Arabidopsis and hybrid poplar.

View Article: PubMed Central - PubMed

Affiliation: Department of Wood Science, University of British Columbia, 2424 Main Mall, Vancouver, BC, V6T 1Z4, Canada.

No MeSH data available.


Related in: MedlinePlus

BRET signal ratio in Arabidopsis hypocotyls from seedlings 7 d after germination using 1 μM coelenterazine at 22 °C indicates a significant (P<0.05) difference in the BRET ratio of RSPS in the Atspp– mutant background and SPSR in the Atsps– mutant background, when compared with the RLUC negative control. P35S::hRLUC·YFP and P35S::hRLUC structures were transformed for a positive control and negative control, respectively.
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Figure 4: BRET signal ratio in Arabidopsis hypocotyls from seedlings 7 d after germination using 1 μM coelenterazine at 22 °C indicates a significant (P<0.05) difference in the BRET ratio of RSPS in the Atspp– mutant background and SPSR in the Atsps– mutant background, when compared with the RLUC negative control. P35S::hRLUC·YFP and P35S::hRLUC structures were transformed for a positive control and negative control, respectively.

Mentions: Following confirmation of YFP expression, a BRET assay was conducted on the double transgenic lines. In a BRET experiment, the extent of energy transfer from RLUC to YFP is measured via a luminometer by filtering the signal luminescence through blue and yellow filters, respectively, and calculating the ratio between yellow and blue signals. Here, the yellow/blue (Y/B) luminescence ratios of 10-day-old light-grown double transgenic seedlings, as well as the negative and positive controls, were calculated. The resultant BRET ratio of the C-tagged RLUC AtSPS in the Atsps– mutant background and the N-tagged RLUC AtSPS in the Atspp– mutant background was significantly higher than that of the RLUC fusion expressed alone as a negative control in the mutant backgrounds (Fig. 4). There was a slight signal discrepancy, of ~0.2, when comparing the interaction in the different mutant backgrounds, with slightly stronger signals in the Atspp– mutant background. According to Subramanian et al. (2006), a BRET signal is considered positive when the yellow to blue ratio is >0.04 compared with the RLUC fusion expressed alone as a negative control. Here, the majority (70%) of 50 pairings of gene products tested showed a ratio >0.04, with some as high as 0.3. These findings suggest a positive protein–protein interaction with these constructs. However, a significant difference in the BRET ratio was not seen in N-tagged RLUC AtSPS in the Atsps– mutant background and the C-tagged RLUC AtSPS in the Atspp– mutant background. While the exact reason for this is unknown, it has been suggested that an unfavourable orientation of the emission dipoles of RLUC and YFP or an excessive distance due to their spatial separation by the larger fused proteins will abolish the BRET signal (Subramanian et al., 2004, 2006). These data taken together with the protein–protein interaction results observed in the yeast two-hybrid assays suggest that there is an active interaction between AtSPS and AtSPP.


Sucrose phosphate synthase and sucrose phosphate phosphatase interact in planta and promote plant growth and biomass accumulation.

Maloney VJ, Park JY, Unda F, Mansfield SD - J. Exp. Bot. (2015)

BRET signal ratio in Arabidopsis hypocotyls from seedlings 7 d after germination using 1 μM coelenterazine at 22 °C indicates a significant (P<0.05) difference in the BRET ratio of RSPS in the Atspp– mutant background and SPSR in the Atsps– mutant background, when compared with the RLUC negative control. P35S::hRLUC·YFP and P35S::hRLUC structures were transformed for a positive control and negative control, respectively.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493782&req=5

Figure 4: BRET signal ratio in Arabidopsis hypocotyls from seedlings 7 d after germination using 1 μM coelenterazine at 22 °C indicates a significant (P<0.05) difference in the BRET ratio of RSPS in the Atspp– mutant background and SPSR in the Atsps– mutant background, when compared with the RLUC negative control. P35S::hRLUC·YFP and P35S::hRLUC structures were transformed for a positive control and negative control, respectively.
Mentions: Following confirmation of YFP expression, a BRET assay was conducted on the double transgenic lines. In a BRET experiment, the extent of energy transfer from RLUC to YFP is measured via a luminometer by filtering the signal luminescence through blue and yellow filters, respectively, and calculating the ratio between yellow and blue signals. Here, the yellow/blue (Y/B) luminescence ratios of 10-day-old light-grown double transgenic seedlings, as well as the negative and positive controls, were calculated. The resultant BRET ratio of the C-tagged RLUC AtSPS in the Atsps– mutant background and the N-tagged RLUC AtSPS in the Atspp– mutant background was significantly higher than that of the RLUC fusion expressed alone as a negative control in the mutant backgrounds (Fig. 4). There was a slight signal discrepancy, of ~0.2, when comparing the interaction in the different mutant backgrounds, with slightly stronger signals in the Atspp– mutant background. According to Subramanian et al. (2006), a BRET signal is considered positive when the yellow to blue ratio is >0.04 compared with the RLUC fusion expressed alone as a negative control. Here, the majority (70%) of 50 pairings of gene products tested showed a ratio >0.04, with some as high as 0.3. These findings suggest a positive protein–protein interaction with these constructs. However, a significant difference in the BRET ratio was not seen in N-tagged RLUC AtSPS in the Atsps– mutant background and the C-tagged RLUC AtSPS in the Atspp– mutant background. While the exact reason for this is unknown, it has been suggested that an unfavourable orientation of the emission dipoles of RLUC and YFP or an excessive distance due to their spatial separation by the larger fused proteins will abolish the BRET signal (Subramanian et al., 2004, 2006). These data taken together with the protein–protein interaction results observed in the yeast two-hybrid assays suggest that there is an active interaction between AtSPS and AtSPP.

Bottom Line: Herein, the formation of an enzyme complex between SPS and SPP was examined, and the results from yeast two-hybrid experiments suggest that there is indeed an association between these proteins.The findings clearly demonstrated that SPS interacts with SPP and that this interaction impacts soluble carbohydrate pools and affects carbon partitioning to starch.Moreover, a fusion construct between the two genes promotes plant growth in both transgenic Arabidopsis and hybrid poplar.

View Article: PubMed Central - PubMed

Affiliation: Department of Wood Science, University of British Columbia, 2424 Main Mall, Vancouver, BC, V6T 1Z4, Canada.

No MeSH data available.


Related in: MedlinePlus