,190-bp fragment containing TGSGP and the associated genomic region. We identified a DNA rearrangement linked to the insertion of TGSGP, which interrupted a gene homologous to AUT1 from S. cerevisiae. In T. b. brucei, this gene is located on chromosome II, ≈ 0.35 Mb from its left-arm telomere32 and is transcribed away from this telomere. The rearrangement occurred in a simple AT-rich region resembling the repeats typically found upstream from telomeric VSG genes.29 Southern blot, PCR, and PFGE analyses indicated that the T. b. gambiense genome is heterozygous for AUT1; it contains AUT1 and a truncated aut1 allele, each present on different chromosome II homologues, as shown by the presence of the PLC marker. The size of the two chromosome II homologues from T. b. gambiense was quite different, 2.7 and 1.1 Mb, respectively. Such size differences are common in trypanosomes, because in the T. b. brucei stock 427 chromosome Ia is 1.85 Mb and chromosome Ib is 3.6 Mb.31 At the current status of the T. b. gambiense genome database (http://www.sanger.ac.uk/Projects/T_b_gambiense/), a sequence containing TGSGP is present (707f06.q1k). This sequence contains 1,057 bp of AT-rich sequence before the ATG codon of TGSGP, but it lacks the junction with the truncated aut1 allele. The main difference between 707f06.q1k (DAL 972) and the corresponding sequence in our LiTat 1.3A stock is a deletion of 98 bp in 707f06.q1k and five other nucleotide changes. At the protein level, the difference between the ORF (403 aa) of the two sequences is only three amino acids, of which only one constitutes a conservative change (data not shown).
As mentioned above, our previous results clearly showed that TGSGP was a telomeric gene located few kbp from a chromosomal end.20 This together with the fact that TGSGP is directly linked to a truncated AUT1 gene (Figure 2, A and B) indicated either: (i) a massive deletion (≈ 0.8 Mb, by comparison to the T. b. brucei’s 1.1-Mb chromosome II) between AUT1 and the TGSGP-containing telomere or (ii) an internal inversion implicating the 3′ region of AUT1 and upstream-linked region (≈ 0.35 Mb). Because the PLC chromosome II marker is still present in the TGSGP-containing chromosome (Figure 3), this suggests that no loss of downstream sequence from aut1 occurred but rather an internal inversion of the left arm of chromosome II (Figure 5). Alternative and more complex explanations would imply loss of chromosome II synteny between T. b. brucei and T. b. gambiense. The recombination target for this inversion was probably the G/A AT-rich region normally found within the ORF of the AUT1 allele, in a sequence coding for eight Asp and five Asn residues. Homologous recombination presumably occurred with a AT-rich barren region commonly found upstream of telomeric VSG genes.29 The recombination, possibly promoted by inverted repeats,33 caused inversion of a 0.35-Mb region without substantial loss of genetic information (except for an AUT1 allele) but with inversion of the direction of transcription (Figure 5). Interestingly, in T. b. brucei, the region downstream from AUT1 (Tb927_02_v4:347536–389002) shows a change in transcription direction, which may explain why such inversion was tolerated. A direct consequence of this inversion was the insertion of a RNA polymerase II-mediated transcription unit near a telomere that could have contained a VSG gene. It was probably this ancestral VSG gene that became TGSGP through evolution and selection of its modern function, or alternatively, it was acquired from the VSG reservoir by a recombination mechanism common to all VSG genes during antigenic variation.34 Actually, in T. b. brucei stock TREU 927/4 (genome project strain), there is a VSG gene highly homologous to TGSGP (81% identity at nucleotide level) (Tb10.v4.0178). It codes for a longer VSG than TGSGP and it may represent a common ancestor for both TGSGP and the LiTat 1.23 VSG. Tb10.v4.0178 is a VSG at one end of chromosome X surrounded by other VSG genes that together may be part of the VSG reservoir. What is compelling is the fact that TGSGP has been maintained in all type 1 T. b. gambiense isolates analyzed to date.19,20 This conservation goes beyond TGSGP and extends to the truncation of one AUT1 allele and inversion of a substantial part of one of the two chromosome II homologues. The region bearing the breakage point in aut1 varied from 3.6 to > 8 kb among 18 different T. b. gambiense isolates, presumably due to the simple nature of this AT-rich region and/or to the lack of coding genes. On the basis of this size variation, three major groups could be distinguished: isolates originating from the Ivory Coast region, from Uganda, and from the West Africa region (Congo, Democratic Republic of Congo, and Cameroon). This finding could be useful to determine the possible geographical origin of T. b. gambiense isolates.
Key to stock origins: year of isolation/original name of isolate. All hosts are human. DRC, Democratic Republic of the Congo. The stocks in bold print are those used in the Northern Blot analysis.
Authors’ addresses: Cecile Felu, Julie Pasture, Etienne Pays, and David Pérez-Morga, Laboratory of Molecular Parasitology, IBMM, Université Libre de Bruxelles, 12, rue des Professeurs Jeener et Brachet, B-6041 Gosselies, Belgium. Telephone: +32-2-650-97 67, Fax: +32-2-650-97 50, E-mail |||EMAIL_ADDRESS||| Acknowledgments: We acknowledge M. Berberof for earlier participation in this project. We are grateful to G. Kanmogne, W. Gibson, P. Büscher, and F. Claes for providing materials.
Financial support: This work was supported by the Belgium Fund for Scientific Research (FRSM) and by the Interuniversity Attraction Poles Program—Belgian Science Policy. C.F. was supported by FRIA.
UK Chart
Remember when the chart was full of £1 PC games that Asda was flogging? Something similar has happened. See, Morrisons had quite a good deal on last week - selected new Xbox 360 and PlayStation 3 games were £25, Wii games £15 and DS games £12. This seems to have had a knock on effect on the chart, with most of the games in the offer shooting up or reappearing. Most notably: Kung Fu Panda back in at #23; Smash Bros Brawl, Off Road, Cooking Guide, Guitar Hero: On Tour, Sports Party, Soul Calibur IV and GTA IV all going up a few places.
The top three has remained the same - Mercenaries 2 on top followed by Tiger Woods 09 and Spore. TNA Impact goes in at #9 despite average review scores, Spore Creatures arrives at #14 in its second week of release, while The Sims 2: Apartment Pets - which apparently sold out on release - is back at #19. Rock Band - which came out on PlayStation 2 and 3 and Wii last week - only manages a poor #30. Also feel sorry for Viva Pinata: Trouble in Paradise, which has slipped from #23 to #40. Too Human is also on the way out dropping from #16 to #38.
There’s a little bit of action in the Xbox 360 chart - Warhammer: Battle March enters at #11, NHL 09 at #15 and Supreme Commander at #25. Ferrari Challenge screeches in at #29 in the Wii chart and something called Spellbound at a lowly #41 in the DS chart.Cedar Grove and Cedar Vale are in the same region to the south of Jimboomba. When the Beaudesert Railway was built in 1888, the station in the area was known as Cedar Pocket.
Much of the land to the west of Cedar Pocket was originally selected by the Markwell family, some of which was later purchased by William Geddes. During the 1887 flood, Geddes lost 20 brood mares with foals, as well as one house with fencing.
In 1872, Geddes’ daughter Susanna married William Henry Couldrey. Couldrey purchased the Ageston sugar plantation on the Logan River in 1868. He was already the owner of the Phoenix and Smithfield mines in Gympie and an active member of the Gympie Community. He employed a manager for Ageston and continued his interests in Gympie. He was the first Chairman of the Gympie Divisional Board.
In mid-1891, Couldrey moved to the Geddes property at Cedar Pocket and began to build a cheese factory. By December 1892, the factory was in full swing with 20 farmers supplying its milk. The factory was located at the intersection of Couldrey Court and Cedar Grove Road.
In later years, the factory was operated by Reg Couldrey, son of William and Susanna Couldrey. In