Journal:

Article Title: A crucial role for the putative Arabidopsis topoisomerase VI in plant growth and development

doi: 10.1073/pnas.152337599

Figure Lengend Snippet: Identification of BIN5 and BIN3 genes. (A) bin5 chromosome walk. The bin5 mutation was mapped to a 46-kb region on BAC F9G14. Complementation constructs B and C, both containing ORF20, rescued the bin5 mutant phenotypes, indicating that ORF20 is mutated in bin5. (B) BIN5/AtSPO11-3 gene and bin5 mutation. BIN5 was identified as AtSPO11-3, one of three Arabidopsis homologs of archaebacterial topoisomerase VI subunit A (Top6A) (23, 25). BIN5 contains two exons and DNA sequencing of bin5 mutant DNA revealed a single base pair addition at nucleotide position +1035 with respect to the putative translation start site (ccg agt gat to ccc gag tga t). A phylogenetic distance tree shows that BIN5 is most closely related to AtSPO11-2 and archaebacterial TOP6A than to nematode, mouse, fly, fungus, and yeast SPO11. Numbers in branch nodes indicate the number of times that the group of sequences to the right of the node occurred of 100 bootstrap replicates. Numbers are only given when 70 or more occurrences were observed. (C) BIN3/AtTop6B gene and mutations. BIN3 was identified by plasmid rescue with a T-DNA-tagged allele, bin3-1. BIN3/AtTOP6B gene contains 18 introns and 19 exons that encode a predicted polypeptide of 670 aa (25). Phylogenetic distance tree shows relationship of BIN3/AtTOP6B, archaebacterial TOP6B, and prokaryotic homologs of Aeropyrum pernix, Archaeoglobus fulgidus, Pyrococcus horikoshii, and Pyrococcus abyssi.

Article Snippet: Genes that are down-regulated by at least 2-fold in both bin3 and bin5 compared with wild-type control were identified using GENESPRING 4.2 (Silicon Genetics, Redwood City, CA) and listed in Table 2, which is published as supporting information on the PNAS web site, www.pnas.org . table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Gene no. Gene Col bin3 bin5 bri1-116 −BL +BL −BL +BL −BL +BL −BL +BL At4g38850 SAUR-AC1 442 2260 161 449 105 476 223 128 At4g38860 SAUR-AC1-like 652 2808 99 834 264 975 285 208 At3g15540 IAA19 980 1958 301 535 372 472 360 438 At2g14900 GASA3 10567 13722 4754 6042 2698 5365 7635 7332 At5g57560 TCH4 19164 26329 14615 15691 12952 16996 5823 5046 At2g26710 BAS1 922 2527 589 1286 954 1150 627 669 At2g31730 Similar to LeER33 1939 2820 1311 1670 1549 1561 1023 1306 At1g65310 Putative xyloglucan endoglucanase 1296 2431 302 394 419 287 274 397 At1g04610 YUCCA3 1372 1639 288 452 532 446 593 653 At5g13870 EXGT-A4 1025 1885 280 267 189 450 637 644 At4g20780 Putative Ca-dependent protein kinase 5508 6829 1223 1225 1077 1265 2532 2130 At2g40610 Putative expansin 3945 7978 615 2767 1011 2070 1178 802 At4g28780 Pro-rich APG like protein 2263 4792 861 2559 531 1995 1373 859 At1g21820 Unknown 2227 4069 1305 1617 1760 1982 1846 1851 At2g32560 Unknown 2534 4380 1235 1817 1516 1519 1366 1769 At2g36220 Unknown 1425 3803 1488 800 1034 1391 1311 1435 At4g09890 Unknown 1497 2725 340 465 388 464 582 657 At4g01950 Unknown 1327 2921 893 1802 946 1465 319 239 Open in a separate window RNA from seedlings treated without or with BL was used for microarray experiments using Affymetrix arabidopsis gene chip .

Techniques: Mutagenesis, Construct, DNA Sequencing, Plasmid Preparation