title: Reconstitution of the Mycobacterium tuberculosis pupylation pathway in Escherichia coli.
abstract: Prokaryotic ubiquitin-like protein (Pup) is a post-translational modifier that attaches to more than 50 proteins in Mycobacteria. Proteasome accessory factor A (PafA) is responsible for Pup conjugation to substrates, but the manner in which proteins are selected for pupylation is unknown. To address this issue, we reconstituted the pupylation of model Mycobacterium proteasome substrates in Escherichia coli, which does not encode Pup or PafA. Surprisingly, Pup and PafA were sufficient to pupylate at least 51 E. coli proteins in addition to the mycobacterial proteins. These data suggest that pupylation signals are intrinsic to targeted proteins and might not require Mycobacterium-specific cofactors for substrate recognition by PafA in vivo.
authors: Department of Microbiology, New York University School of Medicine, 550 First Avenue, MSB 236, New York, New York 10016, USA.
authors: Francisca A
authors: FA
authors: Cerda-Maira
authors: Fiona
authors: F
authors: McAllister
authors: Nadine J
authors: NJ
authors: Bode
authors: Kristin E
authors: KE
authors: Burns
authors: Steven P
authors: SP
authors: Gygi
authors: K Heran
authors: KH
authors: Darwin
mesh_terms: D019732:Amide Synthases; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D014452:Ubiquitins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D014452:Ubiquitins; D046988:Proteasome Endopeptidase Complex; D019732:Amide Synthases; C479921:Paf protein, Mycobacterium tuberculosis