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The small G protein Arl5 contributes to endosome-to-Golgi traffic by aiding the recruitment of the GARP complex to the Golgi.

Rosa-Ferreira C, Christis C, Torres IL, Munro S - Biol Open (2015)

Bottom Line: In addition, in HeLa cells GARP also becomes cytosolic upon depletion of Arl5b.These phenotypes are consistent with a role in endosome-to-Golgi traffic, but are less severe than loss of GARP itself.Thus it appears that Arl5 is one of the factors that directs the recruitment of the GARP complex to the trans-Golgi, and this function is conserved in both flies and humans.

View Article: PubMed Central - PubMed

Affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.

No MeSH data available.


Related in: MedlinePlus

Generation of a  allele for the gene CG7197/Arl5.(A) Schematic illustration of the genomic region containing the Arl5 and its flanking genes. The P element P{RS5} represented was excised imprecisely, generating a  mutant lacking the coding region of the Arl5 gene (CG7197) as well as its 3′UTR and part of the 5′UTR (depicted by the red line below the Arl5 gene). The grey areas indicate untranslated regions (UTR) of the illustrated genes and the blue region represents the coding region of the Arl5 gene. (B) Arl5 is absent from the Arl5KO1 mutant line. Immunoblot of adult fly lysates from OregonR (WT) and w1118; Arl5KO1/Arl5KO1 probed with a rabbit polyclonal antibody against Arl5 and with an anti-β-Actin polyclonal antibody, used as a loading control. (C) Immunoblot of lysates corresponding to a range of fly tissues and to various stages of development were probed with antibodies against Arl5 and Actin (used as a loading control) revealed that Arl5 is widely expressed.
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f01: Generation of a allele for the gene CG7197/Arl5.(A) Schematic illustration of the genomic region containing the Arl5 and its flanking genes. The P element P{RS5} represented was excised imprecisely, generating a mutant lacking the coding region of the Arl5 gene (CG7197) as well as its 3′UTR and part of the 5′UTR (depicted by the red line below the Arl5 gene). The grey areas indicate untranslated regions (UTR) of the illustrated genes and the blue region represents the coding region of the Arl5 gene. (B) Arl5 is absent from the Arl5KO1 mutant line. Immunoblot of adult fly lysates from OregonR (WT) and w1118; Arl5KO1/Arl5KO1 probed with a rabbit polyclonal antibody against Arl5 and with an anti-β-Actin polyclonal antibody, used as a loading control. (C) Immunoblot of lysates corresponding to a range of fly tissues and to various stages of development were probed with antibodies against Arl5 and Actin (used as a loading control) revealed that Arl5 is widely expressed.

Mentions: Unless otherwise stated, fly stocks were maintained at 25C and w1118 was used as the wild-type control. Mutagenesis of the Arl5 gene (CG7197) was done by imprecise excision of the P element P{RS5}5-HA-1500 located at its 5′UTR as illustrated in Fig. 1A. The stock was obtained from Bloomington Stock Centre, confirmed by PCR of the flanking sequences and outcrossed to y1, w1118 for three generations prior to excision, Detection of genomic deletions was then performed by PCR. Cloning and sequencing of the Arl5KO1 allele confirmed the deletion of the entire coding region including the 3′ UTR and part of the 5′ UTR (the 1050 base pairs 8298517–8299566 inclusive replaced by 34 base pairs of uncertain origin).


The small G protein Arl5 contributes to endosome-to-Golgi traffic by aiding the recruitment of the GARP complex to the Golgi.

Rosa-Ferreira C, Christis C, Torres IL, Munro S - Biol Open (2015)

Generation of a  allele for the gene CG7197/Arl5.(A) Schematic illustration of the genomic region containing the Arl5 and its flanking genes. The P element P{RS5} represented was excised imprecisely, generating a  mutant lacking the coding region of the Arl5 gene (CG7197) as well as its 3′UTR and part of the 5′UTR (depicted by the red line below the Arl5 gene). The grey areas indicate untranslated regions (UTR) of the illustrated genes and the blue region represents the coding region of the Arl5 gene. (B) Arl5 is absent from the Arl5KO1 mutant line. Immunoblot of adult fly lysates from OregonR (WT) and w1118; Arl5KO1/Arl5KO1 probed with a rabbit polyclonal antibody against Arl5 and with an anti-β-Actin polyclonal antibody, used as a loading control. (C) Immunoblot of lysates corresponding to a range of fly tissues and to various stages of development were probed with antibodies against Arl5 and Actin (used as a loading control) revealed that Arl5 is widely expressed.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400590&req=5

f01: Generation of a allele for the gene CG7197/Arl5.(A) Schematic illustration of the genomic region containing the Arl5 and its flanking genes. The P element P{RS5} represented was excised imprecisely, generating a mutant lacking the coding region of the Arl5 gene (CG7197) as well as its 3′UTR and part of the 5′UTR (depicted by the red line below the Arl5 gene). The grey areas indicate untranslated regions (UTR) of the illustrated genes and the blue region represents the coding region of the Arl5 gene. (B) Arl5 is absent from the Arl5KO1 mutant line. Immunoblot of adult fly lysates from OregonR (WT) and w1118; Arl5KO1/Arl5KO1 probed with a rabbit polyclonal antibody against Arl5 and with an anti-β-Actin polyclonal antibody, used as a loading control. (C) Immunoblot of lysates corresponding to a range of fly tissues and to various stages of development were probed with antibodies against Arl5 and Actin (used as a loading control) revealed that Arl5 is widely expressed.
Mentions: Unless otherwise stated, fly stocks were maintained at 25C and w1118 was used as the wild-type control. Mutagenesis of the Arl5 gene (CG7197) was done by imprecise excision of the P element P{RS5}5-HA-1500 located at its 5′UTR as illustrated in Fig. 1A. The stock was obtained from Bloomington Stock Centre, confirmed by PCR of the flanking sequences and outcrossed to y1, w1118 for three generations prior to excision, Detection of genomic deletions was then performed by PCR. Cloning and sequencing of the Arl5KO1 allele confirmed the deletion of the entire coding region including the 3′ UTR and part of the 5′ UTR (the 1050 base pairs 8298517–8299566 inclusive replaced by 34 base pairs of uncertain origin).

Bottom Line: In addition, in HeLa cells GARP also becomes cytosolic upon depletion of Arl5b.These phenotypes are consistent with a role in endosome-to-Golgi traffic, but are less severe than loss of GARP itself.Thus it appears that Arl5 is one of the factors that directs the recruitment of the GARP complex to the trans-Golgi, and this function is conserved in both flies and humans.

View Article: PubMed Central - PubMed

Affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.

No MeSH data available.


Related in: MedlinePlus