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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

Transmission electron microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef. A−C, wild type; D−J, the double recessive type from the crossing of EM425 and EM939. A−C depict sections of developing wild-type endosperm at 1 WAF, 2 WAF and 3 WAF, respectively. Corresponding sections from the double recessive type are shown in D−F, G and H, and I and J. I, J: gold particles of 15nm and 5nm indicate the reaction of glutelin and prolamine antibodies, respectively. The square area in J shows the PSV is indicated as enlarged image in Fig. S3F. Arrows in A, D, F, H, I, and J show the PSVs. CW, cell wall; PMB, paramural body. Bars:1 μm in A to F; 2 μm in G to J.
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Figure 3: Transmission electron microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef. A−C, wild type; D−J, the double recessive type from the crossing of EM425 and EM939. A−C depict sections of developing wild-type endosperm at 1 WAF, 2 WAF and 3 WAF, respectively. Corresponding sections from the double recessive type are shown in D−F, G and H, and I and J. I, J: gold particles of 15nm and 5nm indicate the reaction of glutelin and prolamine antibodies, respectively. The square area in J shows the PSV is indicated as enlarged image in Fig. S3F. Arrows in A, D, F, H, I, and J show the PSVs. CW, cell wall; PMB, paramural body. Bars:1 μm in A to F; 2 μm in G to J.

Mentions: To obtain additional insight on the trafficking pathways of glutelin and α-globulin to the PSVs and PMBs in the double recessive type, transmission electron microscopy studies were conducted (Figs 3, 4; Supplementary Fig. S2). At 1 WAF, electron-dense granules and the PMBs were detected in the paramural space created by the invagination of the plasma membrane away from the cell wall in the double recessive type (Fig. 3D, E) as well as in each single mutant (Supplementary Fig. S2B, F) but not in the wild type (Fig. 3A). The extracellular electron-dense granules in the double recessive type were labelled with glutelin antibodies (Supplementary Fig. S3B). This observation together with the reduction of glutelin-containing PSVs in the cytoplasm indicates that proglutelin granules, likely in the form of Golgi-derived dense vesicles, were secreted (Fig. 3D). Some proglutelins were normally transported to PSV in the double recessive type (Fig. 3D, F) as well as in each single mutant (Supplementary Fig. S2A, E). In 2 WAF endosperm, the PMBs in the double recessive type were substantially larger than those observed at 1 WAF (Fig. 3G) and from those present in each single mutant (Supplementary Fig. S2C, G). Unlike the condition seen at 1 WAF, electron-dense granules were not readily observed in the cell wall areas suggesting that these granules were taken up by the PMBs. Similar to that seen for the cell wall-associated electron dense granules, those located in the PMB were labelled with glutelin antibody in the double recessive type (Supplementary Fig. S3D, E).


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)

Transmission electron microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef. A−C, wild type; D−J, the double recessive type from the crossing of EM425 and EM939. A−C depict sections of developing wild-type endosperm at 1 WAF, 2 WAF and 3 WAF, respectively. Corresponding sections from the double recessive type are shown in D−F, G and H, and I and J. I, J: gold particles of 15nm and 5nm indicate the reaction of glutelin and prolamine antibodies, respectively. The square area in J shows the PSV is indicated as enlarged image in Fig. S3F. Arrows in A, D, F, H, I, and J show the PSVs. CW, cell wall; PMB, paramural body. Bars:1 μm in A to F; 2 μm in G to J.
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Related In: Results  -  Collection

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Figure 3: Transmission electron microscopy of the endosperm from the double recessive type of glup4/rab5a and glup6/gef. A−C, wild type; D−J, the double recessive type from the crossing of EM425 and EM939. A−C depict sections of developing wild-type endosperm at 1 WAF, 2 WAF and 3 WAF, respectively. Corresponding sections from the double recessive type are shown in D−F, G and H, and I and J. I, J: gold particles of 15nm and 5nm indicate the reaction of glutelin and prolamine antibodies, respectively. The square area in J shows the PSV is indicated as enlarged image in Fig. S3F. Arrows in A, D, F, H, I, and J show the PSVs. CW, cell wall; PMB, paramural body. Bars:1 μm in A to F; 2 μm in G to J.
Mentions: To obtain additional insight on the trafficking pathways of glutelin and α-globulin to the PSVs and PMBs in the double recessive type, transmission electron microscopy studies were conducted (Figs 3, 4; Supplementary Fig. S2). At 1 WAF, electron-dense granules and the PMBs were detected in the paramural space created by the invagination of the plasma membrane away from the cell wall in the double recessive type (Fig. 3D, E) as well as in each single mutant (Supplementary Fig. S2B, F) but not in the wild type (Fig. 3A). The extracellular electron-dense granules in the double recessive type were labelled with glutelin antibodies (Supplementary Fig. S3B). This observation together with the reduction of glutelin-containing PSVs in the cytoplasm indicates that proglutelin granules, likely in the form of Golgi-derived dense vesicles, were secreted (Fig. 3D). Some proglutelins were normally transported to PSV in the double recessive type (Fig. 3D, F) as well as in each single mutant (Supplementary Fig. S2A, E). In 2 WAF endosperm, the PMBs in the double recessive type were substantially larger than those observed at 1 WAF (Fig. 3G) and from those present in each single mutant (Supplementary Fig. S2C, G). Unlike the condition seen at 1 WAF, electron-dense granules were not readily observed in the cell wall areas suggesting that these granules were taken up by the PMBs. Similar to that seen for the cell wall-associated electron dense granules, those located in the PMB were labelled with glutelin antibody in the double recessive type (Supplementary Fig. S3D, E).

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