Abstract
The symmetry of the phi 29 head-tail connector is controversial: several studies of two-dimensional arrays of the connector have found a 12-fold symmetry, while a recent study of isolated particles has found a 13-fold symmetry. To investigate whether a polymorphism of the structure might explain these different results, electron microscopy and image analysis were used to study both isolated connectors and particles in hexagonally packed arrays. The hexagonally packed arrays have a P1 symmetry, and the connectors displayed 13 subunits both in the arrays and as isolated single particles. While we do not observe a polymorphism between connectors in two-dimensional arrays and as isolated particles, data show that the connectors can exist with either 12 or 13 subunits. A three-dimensional reconstruction of our 13-fold connector was generated by combining an averaged side-view projection with the known symmetry. The structure of rosettes of the connectors formed in the presence of phi 29 prohead RNA (pRNA) was also examined. These rosettes contain five connectors arranged about a single connector in the center, and this arrangement may reflect an essential role of the pRNA in mediating a symmetry mismatch between either a 12- or 13-fold symmetric connector and a putative fivefold symmetric prohead portal vertex into which the connector fits.
MeSH Terms
Bacillus Phages/genetics,ultrastructure
Escherichia coli
Genes, Viral
Microscopy, Electron/methods
Models, Structural
RNA, Viral/ultrastructure
Viral Proteins/ultrastructure
Chemicals
RNA, Viral
Viral Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tsuprun V
Department of Cell Biology and Neuroanatomy, University of Minnesota Medical School, Minneapolis 55455-0303.
Anderson D
Egelman E H
References (29)
29 references, click to expand
-
Harmonic analysis of electron microscope images with rotational symmetry.
J Mol Biol. 1971 Aug 28;60(1):123-30
PMID: 5572100
-
Three-dimensional transformation of capsids associated with genome packaging in a bacterial virus.
J Mol Biol. 1993 May 5;231(1):65-74
PMID: 8496966
-
Gene 20 product of bacteriophage T4 its purification and structure.
J Mol Biol. 1981 Nov 15;152(4):641-62
PMID: 7334518
-
Structure of the head-tail connector of bacteriophage phi 29.
J Mol Biol. 1982 Jan 15;154(2):311-24
PMID: 6804634
-
Structural localization of the proteins of the head to tail connecting region of bacteriophage phi 29.
Virology. 1983 Jan 15;124(1):133-43
PMID: 6401885
-
Overproduction and purification of the connector protein of Bacillus subtilis phage phi 29.
Nucleic Acids Res. 1984 Mar 12;12(5):2351-65
PMID: 6324116
-
Bacteriophage lambda preconnectors. Purification and structure.
J Mol Biol. 1984 Apr 15;174(3):433-47
PMID: 6232391
-
Cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of Bacillus subtilis phage phi 29.
Gene. 1984 Oct;30(1-3):87-98
PMID: 6096227
-
Three-dimensional reconstruction of bacteriophage phi 29 neck particles at 2 X 2 nm resolution.
J Mol Biol. 1985 May 5;183(1):79-88
PMID: 4009722
-
The DNA translocating vertex of dsDNA bacteriophage.
Annu Rev Microbiol. 1985;39:109-29
PMID: 2932996
-
Structural study of tetragonal-ordered aggregates of phage phi 29 necks.
J Ultrastruct Res. 1984 Oct;89(1):79-88
PMID: 6544883
-
Structure of phage phi 29 connector protein assembled in vivo.
Virology. 1985 Mar;141(2):190-200
PMID: 3936270
-
Computer graphic display method for visualizing three-dimensional biological structures.
Science. 1986 May 30;232(4754):1113-5
PMID: 3754654
-
Structure of mitochondrial F1-ATPase studied by electron microscopy and image processing.
Biochim Biophys Acta. 1986 Oct 8;851(3):353-60
PMID: 2875733
-
Bacteriophage T3 gene 8 product oligomer structure.
J Ultrastruct Mol Struct Res. 1986 Feb;94(2):105-13
PMID: 3782924
-
A small viral RNA is required for in vitro packaging of bacteriophage phi 29 DNA.
Science. 1987 May 8;236(4802):690-4
PMID: 3107124
-
Three-dimensional reconstruction of the connector of bacteriophage phi 29 at 1.8 nm resolution.
J Mol Biol. 1986 Dec 20;192(4):853-67
PMID: 3586012
-
Purification and organization of the gene 1 portal protein required for phage P22 DNA packaging.
Biochemistry. 1988 Mar 22;27(6):1849-56
PMID: 3288279
-
Bacteriophage T3 connector: three-dimensional structure and comparison with other viral head-tail connecting regions.
J Mol Biol. 1988 May 5;201(1):91-100
PMID: 3262165
-
Novel second-site suppression of a cold-sensitive defect in phage P22 procapsid assembly.
J Mol Biol. 1990 Dec 5;216(3):701-16
PMID: 2258936
-
Role of RNA in bacteriophage phi 29 DNA packaging.
J Struct Biol. 1990 Jul-Sep;104(1-3):70-4
PMID: 2150913
-
Mutant prohead RNAs in the in vitro packaging of bacteriophage phi 29 DNA-gp3.
J Mol Biol. 1992 Feb 20;223(4):991-8
PMID: 1538407
-
Image reconstruction from cryo-electron micrographs reveals the morphopoietic mechanism in the P2-P4 bacteriophage system.
EMBO J. 1992 Mar;11(3):839-46
PMID: 1547786
-
Three-dimensional structure of T3 connector purified from overexpressing bacteria.
J Mol Biol. 1992 Mar 5;224(1):103-12
PMID: 1548694
-
Three-dimensional reconstruction of single particles embedded in ice.
Ultramicroscopy. 1992 Jan;40(1):33-53
PMID: 1580010
-
Supercoiled DNA wraps around the bacteriophage phi 29 head-tail connector.
Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10479-83
PMID: 1438237
-
Two-dimensional crystals of the molecular chaperone GroEL reveal structural plasticity.
J Mol Biol. 1993 Feb 5;229(3):579-84
PMID: 8094463
-
The portal protein of bacteriophage SPP1: a DNA pump with 13-fold symmetry.
EMBO J. 1993 Apr;12(4):1303-9
PMID: 8467790
-
Use of multivariate statistics in analysing the images of biological macromolecules.
Ultramicroscopy. 1981;6(2):187-94
PMID: 7268930