title: The posttranslational modification cascade to the thiopeptide berninamycin generates linear forms and altered macrocyclic scaffolds.
abstract: Berninamycin is a member of the pyridine-containing thiopeptide class of antibiotics that undergoes massive posttranslational modifications from ribosomally generated preproteins. Berninamycin has a 2-oxazolyl-3-thiazolyl-pyridine core embedded in a 35-atom macrocycle rather than typical trithiazolylpyridine cores embedded in 26-atom and 29-atom peptide macrocycles. We describe the cloning of an 11-gene berninamycin cluster from Streptomyces bernensis UC 5144, its heterologous expression in Streptomyces lividans TK24 and Streptomyces venezuelae ATCC 10712, and detection of variant and incompletely processed scaffolds. Posttranslational maturation in S. lividans of both the wild-type berninamycin prepeptide (BerA) and also a T3A mutant generates macrocyclic compounds as well as linear variants, which have failed to form the pyridine and the macrocycle. Expression of the gene cluster in S. venezuelae generates a variant of the 35-atom skeleton of berninamycin, containing a methyloxazoline in the place of a methyloxazole within the macrocyclic framework.
authors: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors: Steven J
authors: SJ
authors: Malcolmson
authors: Travis S
authors: TS
authors: Young
authors: J Graham
authors: JG
authors: Ruby
authors: Peter
authors: P
authors: Skewes-Cox
authors: Christopher T
authors: CT
authors: Walsh
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D047028:Macrocyclic Compounds / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D048170:Streptomyces lividans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C096599:GE 37468 A; D047028:Macrocyclic Compounds; D010455:Peptides; D010456:Peptides, Cyclic; D011498:Protein Precursors; D013844:Thiazoles; C012574:thiocillin; C011405:berninamycin A
keywords: heterologous expression; linear precursors; prepeptide gene replacement
references: Proc Natl Acad Sci U S A. 2009 Feb 24;106(8):2549-53
references: 19196969
references: Appl Environ Microbiol. 2010 Apr;76(7):2335-44
references: 20154110
references: Chem Biol. 2009 Feb 27;16(2):141-7
references: 19246004
references: J Antibiot (Tokyo). 2002 Jan;55(1):83-91
references: 11918071
references: Chem Rev. 2005 Feb;105(2):685-714
references: 15700961
references: Appl Environ Microbiol. 2010 Nov;76(21):7093-101
references: 20851988
references: Chem Biol. 2012 Dec 21;19(12):1600-10
references: 23261603
references: J Gen Microbiol. 1982 Apr;128(4):875-84
references: 6181185
references: Chem Biol. 2011 May 27;18(5):589-600
references: 21609840
references: J Antibiot (Tokyo). 1994 Dec;47(12):1466-72
references: 7844041
references: J Med Chem. 2012 Aug 9;55(15):6934-41
references: 22812377
references: J Am Chem Soc. 2009 Dec 9;131(48):17563-5
references: 19911780
references: Mol Cell. 2008 Apr 11;30(1):26-38
references: 18406324
references: Appl Environ Microbiol. 2010 Aug;76(15):4969-76
references: 20562278
references: ACS Chem Biol. 2012 Mar 16;7(3):429-42
references: 22206579
references: Nat Prod Rep. 2010 Feb;27(2):153-64
references: 20111801
references: Angew Chem Int Ed Engl. 2009;48(37):6770-3
references: 19536800
references: Biochemistry. 1969 Aug;8(8):3303-8
references: 4980191
references: Nat Prod Rep. 2013 Feb;30(2):218-26
references: 23250571
references: J Am Chem Soc. 2009 Apr 1;131(12):4327-34
references: 19265401
references: J Am Chem Soc. 2010 Sep 8;132(35):12182-4
references: 20707374
references: Nat Prod Rep. 2013 Jan;30(1):108-60
references: 23165928
references: J Biol Chem. 2010 Sep 3;285(36):27525-31
references: 20522549
references: J Mol Biol. 2002 May 24;319(1):27-35
references: 12051934
references: Biochemistry. 2006 Jun 6;45(22):6846-57
references: 16734421
references: ACS Chem Biol. 2013 Mar 15;8(3):473-87
references: 23286465
references: Nat Chem Biol. 2010 Jan;6(1):9-18
references: 20016494
references: J Am Chem Soc. 2010 Jun 2;132(21):7519-27
references: 20455532
references: J Am Chem Soc. 2009 Apr 29;131(16):5946-55
references: 19338336
references: Nature. 1978 Apr 27;272(5656):792-5
references: 643068
references: Biochemistry. 2000 Jan 11;39(1):37-45
references: 10625477
references: J Am Chem Soc. 2010 Nov 24;132(46):16324-6
references: 21047073
references: Nature. 1979 Apr 26;278(5707):859-61
references: 440414
references: Proc Natl Acad Sci U S A. 2011 Aug 9;108(32):13053-8
references: 21788474
references: J Antibiot (Tokyo). 1969 Jan;22(1):12-7
references: 4305864