The genome of bacteriophage phiKZ of Pseudomonas aeruginosa.
Bacteriophage phiKZ is a giant virus that efficiently infects Pseudomonas aeruginosa strains pathogenic to human and, therefore, it is attractive for phage therapy. We present here the complete phiKZ genome sequence and a preliminary analysis of its genome structure. The 280,334 bp genome is a linear, circularly permutated and terminally redundant, A+T-rich double-stranded DNA molecule. The phiKZ DNA has no detectable sequence homology to other viruses and microorganisms, and it does not contain NotI, PstI, SacI, SmaI, XhoI, and XmaIII endonuclease restriction sites. The genome has 306 open reading frames (ORFs) varying in size from 50 to 2237 amino acid residues. According to the orientation of transcription, ORFs are apparently organized into clusters and most have a clockwise direction. The phiKZ genome also encodes six tRNAs specific for Met (AUG), Asn (AAC), Asp (GAC), Leu (TTA), Thr (ACA), and Pro (CCA). A putative promoter sequence containing a TATATTAC block was identified. Most potential stem-loop transcription terminators contain the tetranucleotide UUCG loops. Some genes may be assigned as phage-encoded RNA polymerase subunits. Only 59 phiKZ gene products exhibit similarity to proteins of known function from a diversity of organisms. Most of these conserved gene products, such as dihydrofolate reductase, ribonucleoside diphosphate reductase, thymidylate synthase, thymidylate kinase, and deoxycytidine triphosphate deaminase are involved in nucleotide metabolism. However, no virus-encoded DNA polymerase, DNA replication-associated proteins, or single-stranded DNA-binding protein were found based on amino acid homology, and they may therefore be strongly divergent from known homologous proteins. Fifteen phiKZ gene products show homology to proteins of pathogenic organisms, including Mycobacterium tuberculosis, Haemophilus influenzae, Listeria sp., Rickettsia prowazakeri, and Vibrio cholerae that must be considered before using this phage as a therapeutic agent. The phiKZ coat contains at least 40 polypeptides, and several proteins are cleaved during virus assembly in a way similar to phage T4. Eleven phiKZ-encoded polypeptides are related to proteins of other bacteriphages that infect a variety of hosts. Among these are four gene products that contain a putative intron-encoded endonuclease harboring the H-N-H motif common to many double-stranded DNA phages. These observations provide evidence that phages infecting diverse hosts have had access to a common genetic pool. However, limited homology on the DNA and protein levels indicates that bacteriophage phiKZ represents an evolutionary distinctive branch of the Myoviridae family.
Mesyanzhinov
Vadim V
VV
Laboratory of Gene Technology, Katholieke Universiteit Leuven, Kardinaal Mercierlaan 92, B-3001 Leuven, Belgium. vvm@bch.ru
Robben
Johan
J
Grymonprez
Barbara
B
Kostyuchenko
Victor A
VA
Bourkaltseva
Maria V
MV
Sykilinda
Nina N
NN
Krylov
Victor N
VN
Volckaert
Guido
G
D000595:Amino Acid Sequence; D001482:Base Composition; D001483:Base Sequence; D004720:Endonucleases / Q000235:genetics; D019143:Evolution, Molecular; D005814:Genes, Viral / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D016679:Genome, Viral; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D017105:Pseudomonas Phages / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D011550:Pseudomonas aeruginosa / Q000821:virology; D012343:RNA, Transfer / Q000235:genetics; D017422:Sequence Analysis, DNA; D013728:Terminator Regions, Genetic / Q000235:genetics; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics
D014764:Viral Proteins; D012343:RNA, Transfer; D004720:Endonucleases