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Guanine nucleotide exchange factor 2 for Rab5 proteins coordinated with GLUP6/GEF regulates the intracellular transport of the proglutelin from the Golgi apparatus to the protein storage vacuole in rice endosperm.

Wen L, Fukuda M, Sunada M, Ishino S, Ishino Y, Okita TW, Ogawa M, Ueda T, Kumamaru T - J. Exp. Bot. (2015)

Bottom Line: Like GLUP6/GEF, Rab5-GEF2 was capable of activating Rab5a and two other Rab5 isoforms in in vitro GTP/GDP exchange assays.GEF proteins consist of the helical bundle (HB) domain at the N-terminus, Vps9 domain, and a C-terminal region.By the deletion analysis of GEFs, the HB domain was found essential for the activation of Rab5 proteins.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan Tobacco Research Institute, Chinese Academy of Agricultural Science, Qingdao 266101, China.

No MeSH data available.


Related in: MedlinePlus

SDS-PAGE of seed storage proteins in the double recessive type of glup4/rab5a and glup6/gef. (A) CBB staining of the gel. (B) Ratio of proglutelin to sum of glutelin subunits. This value was calculated from densitometry of image A. (C) The floury endosperm of mutant lines. Lane 1, the wild type, Taichung65; lane 2, the wild type, Kinmaze; lane 3, glup4/rab5a, EM425; lane 4, glup4/rab5a, EM956; lane 5, glup6/gef, EM939; lane 6, double recessive type from the crossing of EM425 and EM939; lane 7, double recessive type from the crossing of EM939 and EM956.
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Figure 1: SDS-PAGE of seed storage proteins in the double recessive type of glup4/rab5a and glup6/gef. (A) CBB staining of the gel. (B) Ratio of proglutelin to sum of glutelin subunits. This value was calculated from densitometry of image A. (C) The floury endosperm of mutant lines. Lane 1, the wild type, Taichung65; lane 2, the wild type, Kinmaze; lane 3, glup4/rab5a, EM425; lane 4, glup4/rab5a, EM956; lane 5, glup6/gef, EM939; lane 6, double recessive type from the crossing of EM425 and EM939; lane 7, double recessive type from the crossing of EM939 and EM956.

Mentions: The self-pollinated seeds of the double recessive type were then analysed by SDS-PAGE (Fig. 1A). The levels of proglutelin and glutelin subunits in the double recessive type were substantially elevated and reduced, respectively, compared with each parent line (Fig. 1A). This view is readily evident by analysis of the ratio of proglutelin to glutelin subunits where the double recessive type has a much higher ratio than that seen for each parent line (Fig. 1B). In addition, the grain morphology of the double recessive type exhibited a more severe floury grain phenotype than each parent line (Fig 1C). These results suggest that the glup4 and glup6 mutations interact synergistically to elevate proglutelin and, in turn, reduce glutelin subunits as well as mediate a more severe chalky grain phenotype.


Guanine nucleotide exchange factor 2 for Rab5 proteins coordinated with GLUP6/GEF regulates the intracellular transport of the proglutelin from the Golgi apparatus to the protein storage vacuole in rice endosperm.

Wen L, Fukuda M, Sunada M, Ishino S, Ishino Y, Okita TW, Ogawa M, Ueda T, Kumamaru T - J. Exp. Bot. (2015)

SDS-PAGE of seed storage proteins in the double recessive type of glup4/rab5a and glup6/gef. (A) CBB staining of the gel. (B) Ratio of proglutelin to sum of glutelin subunits. This value was calculated from densitometry of image A. (C) The floury endosperm of mutant lines. Lane 1, the wild type, Taichung65; lane 2, the wild type, Kinmaze; lane 3, glup4/rab5a, EM425; lane 4, glup4/rab5a, EM956; lane 5, glup6/gef, EM939; lane 6, double recessive type from the crossing of EM425 and EM939; lane 7, double recessive type from the crossing of EM939 and EM956.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588877&req=5

Figure 1: SDS-PAGE of seed storage proteins in the double recessive type of glup4/rab5a and glup6/gef. (A) CBB staining of the gel. (B) Ratio of proglutelin to sum of glutelin subunits. This value was calculated from densitometry of image A. (C) The floury endosperm of mutant lines. Lane 1, the wild type, Taichung65; lane 2, the wild type, Kinmaze; lane 3, glup4/rab5a, EM425; lane 4, glup4/rab5a, EM956; lane 5, glup6/gef, EM939; lane 6, double recessive type from the crossing of EM425 and EM939; lane 7, double recessive type from the crossing of EM939 and EM956.
Mentions: The self-pollinated seeds of the double recessive type were then analysed by SDS-PAGE (Fig. 1A). The levels of proglutelin and glutelin subunits in the double recessive type were substantially elevated and reduced, respectively, compared with each parent line (Fig. 1A). This view is readily evident by analysis of the ratio of proglutelin to glutelin subunits where the double recessive type has a much higher ratio than that seen for each parent line (Fig. 1B). In addition, the grain morphology of the double recessive type exhibited a more severe floury grain phenotype than each parent line (Fig 1C). These results suggest that the glup4 and glup6 mutations interact synergistically to elevate proglutelin and, in turn, reduce glutelin subunits as well as mediate a more severe chalky grain phenotype.

Bottom Line: Like GLUP6/GEF, Rab5-GEF2 was capable of activating Rab5a and two other Rab5 isoforms in in vitro GTP/GDP exchange assays.GEF proteins consist of the helical bundle (HB) domain at the N-terminus, Vps9 domain, and a C-terminal region.By the deletion analysis of GEFs, the HB domain was found essential for the activation of Rab5 proteins.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan Tobacco Research Institute, Chinese Academy of Agricultural Science, Qingdao 266101, China.

No MeSH data available.


Related in: MedlinePlus