The holin of bacteriophage lambda forms rings with large diameter.
Holins control the length of the infection cycle of tailed phages (the Caudovirales) by oligomerizing to form lethal holes in the cytoplasmic membrane at a time dictated by their primary structure. Nothing is currently known about the physical basis of their oligomerization or the structure of the oligomers formed by any known holin. Here we use electron microscopy and single-particle analysis to characterize structures formed by the bacteriophage lambda holin (S105) in vitro. In non-ionic or mild zwitterionic detergents, purified S105, but not the lysis-defective variant S105A52V, forms rings of at least two size classes, the most common having inner and outer diameters of 8.5 and 23 nm respectively, and containing approximately 72 S105 monomers. The height of these rings, 4 nm, closely matches the thickness of the lipid bilayer. The central channel is of unprecedented size for channels formed by integral membrane proteins, consistent with the non-specific nature of holin-mediated membrane permeabilization. S105 present in detergent-solubilized rings and in inverted membrane vesicles showed similar sensitivities to proteolysis and cysteine-specific modification, suggesting that the rings are representative of the lethal holes formed by S105 to terminate the infection cycle and initiate lysis.
Savva
Christos G
CG
Dewey
Jill S
JS
Deaton
John
J
White
Rebecca L
RL
Struck
Douglas K
DK
Holzenburg
Andreas
A
Young
Rye
R
D000595:Amino Acid Sequence; D020285:Cryoelectron Microscopy; D003902:Detergents / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*
D003902:Detergents; D008565:Membrane Proteins; C108624:S holin, bacteriophage lambda; D014764:Viral Proteins; D010447:Peptide Hydrolases
J Bacteriol. 1999 May;181(9):2922-9
10217787
Biophys J. 2004 Dec;87(6):4075-86
15465852
EMBO J. 1990 Apr;9(4):981-9
2138979
J Comput Chem. 2004 Oct;25(13):1605-12
15264254
Biophys J. 2005 Mar;88(3):1778-98
15738466
Biochemistry. 2003 Mar 25;42(11):3180-8
12641449
J Mol Biol. 1996 Jul 19;260(3):289-98
8757792
FEBS Lett. 1999 Sep 24;458(3):271-7
10570923
J Mol Biol. 1988 Jan 5;199(1):95-105
2965249
J Mol Biol. 1998 Apr 17;277(5):1033-46
9571020
Genetics. 2006 Jan;172(1):17-26
16219778
Mol Gen Genet. 1981;184(1):111-4
6460914
J Mol Biol. 2004 Feb 13;336(2):453-60
14757057
Protein Sci. 2004 Jul;13(7):1778-86
15215521
J Bacteriol. 1998 May;180(9):2531-40
9573208
J Struct Biol. 1999 Dec 1;128(1):82-97
10600563
Annu Rev Microbiol. 2000;54:799-825
11018145
J Bacteriol. 2000 Nov;182(21):6082-90
11029428
Biochemistry. 1968 Jun;7(6):2143-52
4873173
J Mol Biol. 1983 May 15;166(2):211-7
6854644
Microbiology (Reading). 2008 Jun;154(Pt 6):1710-1718
18524925
Cell. 2005 Apr 22;121(2):247-56
15851031
J Struct Biol. 1996 Jan-Feb;116(1):17-24
8742718
Mol Microbiol. 1999 Aug;33(3):569-82
10417647
Eur J Biochem. 1971 Jan 1;18(1):1-9
5540512
Proc Natl Acad Sci U S A. 2001 Jul 31;98(16):9348-52
11459934
J Biol Chem. 2000 Jan 14;275(2):769-76
10625606
J Bacteriol. 2003 Feb;185(3):779-87
12533453
Mol Microbiol. 1994 Aug;13(3):495-504
7997166
J Mol Biol. 1998 Feb 13;276(1):151-64
9514719
J Bacteriol. 1998 Aug;180(16):4199-211
9696770
J Bacteriol. 1995 Jun;177(11):3283-94
7768829
J Bacteriol. 1990 Feb;172(2):912-21
2137120
J Bacteriol. 1983 Jul;155(1):381-90
6408064