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Autophagy is a pathway by which cytoplasmic constituents, including
organelles and intracellular pathogens, are sequestred in a doublemembrane-bound
autophagosome and delivered to the lysosome for
degradation. The role of autophagy is to eliminate unwanted constituents
from the cell and recycle cytoplasmic material allowing the cell to maintain
macromolecular synthesis and energy homeostatis during starvation and
other stressful conditions. The autophagy pathway proceeds through a
series of stages. It starts with the nucleation of the autophagic vesicle
followed by the elongation and closure of the autophagosome membrane
to envelop cytoplasmic constituents. Then the autophagosome docks with
the lysosome leading to the breakdown of the inner membrane and the
subsequent exposure of the sequestred cytoplasmic material to lysosomal
hydrolases1, 2.
The autophagy pathway requires the concerted action of evolutionarily
conserved genes. Vesicle nucleation depends on a class III
phosphatidylinositol-3-OH kinase (PI(3)K) complex formed by Beclin 1,
Vps34 and other proteins. Atg7 participates in two ubiquitin-like conjugation
pathways, conjugation of Atg5 to Atg12 and conversion of LC3 to its
phosphatidylethanolamine (PE)-conjugated LC3-II form. The Atg5-Atg12
conjugate forms a large complex with Atg16L1. Both conjugation systems
are required for the generation of the autophagosome.
Autophagy plays a key role in the prevention of aging, cancer and
neurodegeneration but also in the innate immune system against intracellular
Autophagy Genes
Description
Autophagy genes are provided as open reading frames (coding sequences)
from the Start codon to the Stop codon. They are cloned in the pUNO
plasmid (see page 58) downstream of the EF-1a/HTLV promoter. pUNO
plasmids are selectable in E. coli and mammalian cells with blasticidin.
Contents and Storage
Each pUNO plasmid is provided as a lyophilized transformed E. coli strain
on a paper disk. Transformed strains are shipped at room temperature and
should be stored at -20°C. They are provided with 4 pouches of E. coli
Fast-Media ® Blas (2 TB and 2 Agar). For more information about
Fast-Media ® , see pages 44-45.
Autophagy and TLRs
pathogens. Autophagy has been shown to interact with pattern recognition
receptors (PRRs), such as the Toll-like receptors (TLRs). It was recently
reported that the autophagic machinery can deliver pathogen-associated
molecular patterns (PAMPs) to endosomal TLRs 3 , suggesting that autophagy
enhances TLR recognition of PAMPs. Conversely, TLRs have been shown to
promote autophagy. Several groups have reported the induction of
autophagy by signaling through TLR4, TLR7, TLR3, TLR2 and TLR5 4-6 . TLRinduced
autophagy appears to depend on MyD88 and TRIF. Both adapters
trigger autophagy through a direct interaction with Beclin 1 (Shi). Autophagy
seems also to participate in the regulation of inflammation. Macrophages
from mice deficient for Atg16L1 produce more of the proinflammatory
cytokines IL-1b and IL-18 after endotoxin stimulation of TLR4 7 . These data
demonstrate that induction of autophagy and TLR signaling are connected.
1. Levine B. & Kroemer G., 2008. Autophagy in the pathogenesis of disease. Cell.
132(1):27-42. Review. 2. Mizushima N. et al., 2008. Autophagy fights disease through
cellular self-digestion. Nature. 451(7182):1069-75. Review. 3. Lee HK. et al., 2007.
Autophagy-dependent viral recognition by plasmacytoid dendritic cells. Science.
315(5817):1398-401. 4. Xu Y. et al., 2007. Toll-like receptor 4 is a sensor for autophagy
associated with innate immunity. Immunity. 27(1):135-44. 5. Delgado MA,. et al., 2008.
Toll-like receptors control autophagy. EMBO J. 27(7):1110-21. 6. Shi CS. &Kehrl JH.,
2008. MyD88 and Trif target Beclin 1 to trigger autophagy in macrophages. J Biol Chem.
283(48):33175-82. 7. Saitoh T. et al., 2008. Loss of the autophagy protein Atg16L1
enhances endotoxin-induced IL-1beta production. Nature. 2456(7219):264-8.
PRODUCT QTY
www.invivogen.com/autophagy
CAT. CODE
(Human)
CAT. CODE
(Mouse)
pUNO1-ATG3 E. coli disk puno1-hatg3 puno1-matg3
pUNO1-ATG4A E. coli disk puno1-hatg4a puno1-matg4a
pUNO1-ATG4B E. coli disk puno1-hatg4b puno1-matg4b
pUNO1-ATG5 E. coli disk puno1-hatg5 puno1-matg5
pUNO1-ATG7 E. coli disk puno1-hatg7 puno1-matg7
pUNO1-ATG10 E. coli disk puno1-hatg10 puno1-matg10
pUNO1-ATG16L1 E. coli disk puno1-hatg16l1 puno1-matg16l1
pUNO1-BECLIN1 E. coli disk puno1-hbecn1 puno1-mbecn1
pUNO1-LC3A E. coli disk puno1-hlc3a puno1-mlc3a
pUNO1-LC3B E. coli disk puno1-hlc3b puno1-mlc3b
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INNATE IMMUNITY 3