A novel application of gene arrays: Escherichia coli array provides insight into the biology of the obligate endosymbiont of tsetse flies

Symbiotic associations with microorganisms are pivotal in many insects. Yet, the functional roles of obligate symbionts have been difficult to study because it has not been possible to cultivate these organisms in vitro. The medically important tsetse fly (Diptera: Glossinidae) relies on its obligate endosymbiont, Wigglesworthia glossinidia, a member of the Enterobacteriaceae, closely related to Escherichia coli, for fertility and possibly nutrition. We show here that the intracellular Wigglesworthia has a reduced genome size smaller than 770 kb. In an attempt to understand the composition of its genome, we used the gene arrays developed for E. coli. We were able to identify 650 orthologous genes in Wigglesworthia corresponding to ≈85% of its genome. The arrays were also applied for expression analysis using Wigglesworthia cDNA and 61 gene products were detected, presumably coding for some of its most abundant products. Overall, genes involved in cell processes, DNA replication, transcription, and translation were found largely retained in the small genome of Wigglesworthia. In addition, genes coding for transport proteins, chaperones, biosynthesis of cofactors, and some amino acids were found to comprise a significant portion, suggesting an important role for these proteins in its symbiotic life. Based on its expression profile, we predict that Wigglesworthia may be a facultative anaerobic organism that utilizes ammonia as its major source of nitrogen. We present an application of E. coli gene arrays to obtain broad genome information for a closely related organism in the absence of complete genome sequence data.

[1]  N. Moran,et al.  Cospeciation of Psyllids and Their Primary Prokaryotic Endosymbionts , 2000, Applied and Environmental Microbiology.

[2]  N. Moran,et al.  Genetics, physiology, and evolutionary relationships of the genus Buchnera: intracellular symbionts of aphids. , 1995, Annual review of microbiology.

[3]  S. Aksoy,et al.  Wigglesworthia gen. nov. and Wigglesworthia glossinidia sp. nov., taxa consisting of the mycetocyte-associated, primary endosymbionts of tsetse flies. , 1995, International journal of systematic bacteriology.

[4]  D. Denlinger,et al.  Secretory discharge and microflora of milk gland in tsetse flies , 1974, Nature.

[5]  N. Moran,et al.  Decoupling of Genome Size and Sequence Divergence in a Symbiotic Bacterium , 2000, Journal of bacteriology.

[6]  R. Fani,et al.  Flavobacteria as intracellular symbionts in cockroaches , 1994, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[7]  Serap Aksoy,et al.  Concordant Evolution of a Symbiont with Its Host Insect Species: Molecular Phylogeny of Genus Glossina and Its Bacteriome-Associated Endosymbiont, Wigglesworthia glossinidia , 1999, Journal of Molecular Evolution.

[8]  S. K. Moloo An artificial feeding technique for Glossina , 1971, Parasitology.

[9]  M. Munson,et al.  Buchnera gen. nov. and Buchnera aphidicola sp. nov., a Taxon Consisting of the Mycetocyte-Associated, Primary Endosymbionts of Aphids , 1991 .

[10]  H. Charles,et al.  Physical and Genetic Map of the Genome of Buchnera, the Primary Endosymbiont of the Pea Aphid Acyrthosiphon pisum , 1999, Journal of Molecular Evolution.

[11]  H. Ochman,et al.  Evolutionary dynamics of full genome content in Escherichia coli , 2000, The EMBO journal.

[12]  S. K. Moloo Aspects of the nutrition of adult female Glossina morsitans during pregnancy. , 1976, Journal of insect physiology.

[13]  E. Bursell Nitrogenous waste products of the tsetse fly, Glossina morsitans. , 1965, Journal of insect physiology.

[14]  A. Khodursky,et al.  Nitrogen regulatory protein C-controlled genes of Escherichia coli: scavenging as a defense against nitrogen limitation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[15]  V. Wigglesworth Digestion in the Tsetse-Fly: A Study of Structure and Function , 1929, Parasitology.

[16]  I. Maudlin,et al.  Sodalis gen. nov. and Sodalis glossinidius sp. nov., a microaerophilic secondary endosymbiont of the tsetse fly Glossina morsitans morsitans. , 1999, International journal of systematic bacteriology.

[17]  D. Saunders,et al.  Letter: The production of "symbiont-free" Glossina morsitans and an associated loss of female fertility. , 1973, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[18]  R. Hess,et al.  The occurrence of intracellular rickettsia-like organisms in the tsetse flies, Glossina morsitans, G. fuscipes, G. brevipalpis and G. pallidipes. , 1974, Acta tropica.

[19]  R. Fleischmann,et al.  The Minimal Gene Complement of Mycoplasma genitalium , 1995, Science.

[20]  N. Moran,et al.  Phylogenetic relationships of the endosymbionts of mealybugs (Homoptera: Pseudococcidae) based on 16S rDNA sequences. , 1992, Molecular phylogenetics and evolution.

[21]  S. Aksoy Tsetse--A haven for microorganisms. , 2000, Parasitology today.

[22]  N. Moran Accelerated evolution and Muller's rachet in endosymbiotic bacteria. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[23]  N. Moran,et al.  Lifestyle evolution in symbiotic bacteria: insights from genomics. , 2000, Trends in ecology & evolution.

[24]  S. Aksoy,et al.  Tissue tropism, transmission and expression of foreign genes in vivo in midgut symbionts of tsetse flies , 1999, Insect molecular biology.

[25]  W. Goebel,et al.  Intracellular endosymbiotic bacteria of Camponotus species (carpenter ants): systematics, evolution and ultrastructural characterization , 1996, Molecular microbiology.

[26]  M. Winkler,et al.  An Escherichia coli K-12 tktA tktB mutant deficient in transketolase activity requires pyridoxine (vitamin B6) as well as the aromatic amino acids and vitamins for growth , 1994, Journal of Bacteriology.

[27]  A. Chow,et al.  Mycetome endosymbionts of tsetse flies constitute a distinct lineage related to Enterobacteriaceae , 1995, Insect molecular biology.

[28]  R. Sinden,et al.  The micro-organisms of tsetse flies. , 1975, Acta tropica.

[29]  S. Aksoy Molecular analysis of the endosymbionts of tsetse flies: 16S rDNA locus and over‐expression of a chaperonin , 1995, Insect molecular biology.

[30]  S. Aksoy,et al.  Phylogeny and potential transmission routes of midgut‐associated endosymbionts of tsetse (Diptera: Glossinidae) , 1997, Insect molecular biology (Print).

[31]  A. Douglas,et al.  Synthesis of the essential amino acid tryptophan in the pea aphid (Acyrthosiphon pisum) symbiosis , 1992 .

[32]  N. Moran,et al.  Bacterial endosymbionts in animals. , 2000, Current opinion in microbiology.

[33]  A. Troesch,et al.  Mycobacterium Species Identification and Rifampin Resistance Testing with High-Density DNA Probe Arrays , 1999, Journal of Clinical Microbiology.

[34]  M. Hattori,et al.  Genome sequence of the endocellular bacterial symbiont of aphids Buchnera sp. APS , 2000, Nature.

[35]  S. K. Moloo,et al.  Comparative study on Rickettsia-like organisms in the midgut epithelial cells of different Glossina species , 1991, Parasitology.