RESUMO
Membrane transport carriers fuse with target membranes through engagement of cognate vSNAREs and tSNAREs on each membrane. How vSNAREs are sorted into transport carriers is incompletely understood. Here we show that VAMP7, the vSNARE for fusing endosome-derived tubular transport carriers with maturing melanosomes in melanocytes, is sorted into transport carriers in complex with the tSNARE component STX13. Sorting requires either recognition of VAMP7 by the AP-3δ subunit of AP-3 or of STX13 by the pallidin subunit of BLOC-1, but not both. Consequently, melanocytes expressing both AP-3δ and pallidin variants that cannot bind their respective SNARE proteins are hypopigmented and fail to sort BLOC-1-dependent cargo, STX13, or VAMP7 into transport carriers. However, SNARE binding does not influence BLOC-1 function in generating tubular transport carriers. These data reveal a novel mechanism of vSNARE sorting by recognition of redundant sorting determinants on a SNARE complex by an AP-3-BLOC-1 super-complex.
Assuntos
Complexo 3 de Proteínas Adaptadoras/genética , Subunidades delta do Complexo de Proteínas Adaptadoras/genética , Proteínas do Tecido Nervoso/genética , Proteínas Qa-SNARE/genética , Proteínas R-SNARE/genética , Endossomos/genética , Humanos , Melanócitos/metabolismo , Melanossomas/genética , Transporte Proteico/genéticaRESUMO
The study of skin color in cattle holds both economic and scientific interest. Several ocular diseases of cattle have been associated with low pigmentation of the eyelids, including ocular squamous cell carcinoma and infectious keratoconjunctivitis, the two most common ocular diseases affecting cattle production. Although low eyelid pigmentation is a well-known risk factor for various ocular diseases, the genetic and biological basis of this relationship is largely unknown. We investigated the transcriptome of eyelid skin in Hereford cattle using RNA-sequencing technology. Two contrasting groups were evaluated: steers that were completely pigmented and steers with no pigmentation in both eyelids. Most of the up-regulated genes in pigmented samples are directly implicated in melanogenesis and melanosome development, whereas up-regulated genes in non-pigmented samples are implicated in cancer development and the immune system, among other functions. Interestingly, network analysis comparing pigmented vs. non-pigmented samples revealed significant differences in the co-expression patterns of genes related to melanosome, pigmentation and defense response to bacteria, showing higher gene activity, greater co-expression patterns and tighter co-regulation mechanisms in pigmented samples. Overall, our findings indicate that bovine eyelid pigmentation depends on the expression of many genes involved not only in pigmentation and melanosome function but also related to inflammatory response, infection and tumoral pathways.