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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

Immunofluorescence microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef at 2 WAF. A−C, wild type; D−F, glup4/rab5a; G to I, glup6/gef; J−L, double recessive type from the crossing of EM425 and EM939. B, E, H, and K are the enlarged images of the square areas in A, D, G, and J, respectively. A, B, D, E, G, H, J, and K: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of prolamine and glutelin antibodies, respectively. C, F, I, and L: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of α-globulin and glutelin antibodies, respectively. Arrows indicates the paramural bodies (PMBs). Bars, 10 μm.
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Figure 2: Immunofluorescence microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef at 2 WAF. A−C, wild type; D−F, glup4/rab5a; G to I, glup6/gef; J−L, double recessive type from the crossing of EM425 and EM939. B, E, H, and K are the enlarged images of the square areas in A, D, G, and J, respectively. A, B, D, E, G, H, J, and K: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of prolamine and glutelin antibodies, respectively. C, F, I, and L: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of α-globulin and glutelin antibodies, respectively. Arrows indicates the paramural bodies (PMBs). Bars, 10 μm.

Mentions: In order to elucidate the influence of the combined glup4/rab5a and glup6/gef mutations on intracellular transport of proglutelins, protein bodies in the endosperm of the single and double recessive type were analysed by immunofluorescence microscopy (Fig. 2, Supplementary Fig. S1). In the wild-type endosperm, protein bodies containing prolamines (protein body type I: PB-I) and those containing glutelins and α-globulins (protein storage vacuole: PSV) increased in size and number as the seed develops (Fig. 2A−C, Supplementary Fig. S1A−C). In each mutant type, prolamine containing PB-I were estimated to be similar in size and number to that seen for the wild type (Fig. 2B, E, H, K). In contrast, glutelin-containing PSVs were smaller and fewer in number in each mutant type (Fig. 2E, H, K) compared with the wild type (Fig. 2B). The severity of this condition varied among the mutant types, with glup4/rab5a showing a moderate reduction in size and number of PSVs (Fig. 2D, E) followed by glup6/gef (Fig. 2G, H) while the double recessive type showed the largest reduction in number of PSVs (Fig. 2J, K). Nearly all of the synthesized α-globulin was packaged in paramural bodies (PMBs) in the double recessive type (Fig. 2L).


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)

Immunofluorescence microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef at 2 WAF. A−C, wild type; D−F, glup4/rab5a; G to I, glup6/gef; J−L, double recessive type from the crossing of EM425 and EM939. B, E, H, and K are the enlarged images of the square areas in A, D, G, and J, respectively. A, B, D, E, G, H, J, and K: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of prolamine and glutelin antibodies, respectively. C, F, I, and L: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of α-globulin and glutelin antibodies, respectively. Arrows indicates the paramural bodies (PMBs). Bars, 10 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 2: Immunofluorescence microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef at 2 WAF. A−C, wild type; D−F, glup4/rab5a; G to I, glup6/gef; J−L, double recessive type from the crossing of EM425 and EM939. B, E, H, and K are the enlarged images of the square areas in A, D, G, and J, respectively. A, B, D, E, G, H, J, and K: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of prolamine and glutelin antibodies, respectively. C, F, I, and L: secondary antibodies labelled with rhodamine (magenta) and fluorescein isothiocyanate (FITC, green) were used to visualize the reaction of α-globulin and glutelin antibodies, respectively. Arrows indicates the paramural bodies (PMBs). Bars, 10 μm.
Mentions: In order to elucidate the influence of the combined glup4/rab5a and glup6/gef mutations on intracellular transport of proglutelins, protein bodies in the endosperm of the single and double recessive type were analysed by immunofluorescence microscopy (Fig. 2, Supplementary Fig. S1). In the wild-type endosperm, protein bodies containing prolamines (protein body type I: PB-I) and those containing glutelins and α-globulins (protein storage vacuole: PSV) increased in size and number as the seed develops (Fig. 2A−C, Supplementary Fig. S1A−C). In each mutant type, prolamine containing PB-I were estimated to be similar in size and number to that seen for the wild type (Fig. 2B, E, H, K). In contrast, glutelin-containing PSVs were smaller and fewer in number in each mutant type (Fig. 2E, H, K) compared with the wild type (Fig. 2B). The severity of this condition varied among the mutant types, with glup4/rab5a showing a moderate reduction in size and number of PSVs (Fig. 2D, E) followed by glup6/gef (Fig. 2G, H) while the double recessive type showed the largest reduction in number of PSVs (Fig. 2J, K). Nearly all of the synthesized α-globulin was packaged in paramural bodies (PMBs) in the double recessive type (Fig. 2L).

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