Fragment-Based Discovery of Low-Micromolar ATAD2 Bromodomain Inhibitors.

Overexpression of ATAD2 (ATPase family, AAA domain containing 2) has been linked to disease severity and progression in a wide range of cancers, and is implicated in the regulation of several drivers of cancer growth. Little is known of the dependence of these effects upon the ATAD2 bromodomain, which has been categorized as among the least tractable of its class. The absence of any potent, selective inhibitors limits clear understanding of the therapeutic potential of the bromodomain. Here, we describe the discovery of a hit from a fragment-based targeted array. Optimization of this produced the first known micromolar inhibitors of the ATAD2 bromodomain.

Tumebacillus lipolyticus sp. nov., isolated from river water.

An aerobic, endospore-forming, alkali-tolerant, Gram-stain-positive, non-motile, rod-shaped bacterium, designated strain NIO-S10T, was isolated from a surface water sample collected from the Godavari River, Kapileswarapuram, India. Colonies on nutrient agar were circular, 3-4 mm in diameter, creamish and raised after incubation for 36 h at 37 °C. Growth occurred at 20-40 °C, at pH 6-9 and in the presence of 0-2 % (w/v) NaCl. Strain NIO-S10T was positive for oxidase, caseinase, DNase, gelatinase, lipase and urease activities, and negative for catalase, aesculinase, amylase and cellulase activities. The fatty acids were dominated by branched and saturated fatty acids, with a high abundance of iso-C15 : 0, anteiso-C15 : 0 and C18 : 0.The cell-wall peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid, and MK-7 was the major menaquinone. The DNA G+C content of strain NIO-S10T was 54.4 mol%. A blast sequence similarity search based on 16S rRNA gene sequences indicated that Tumebacillus ginsengisoli Gsoil 1105T was the nearest phylogenetic neighbour to strain NIO-S10T, with a pairwise sequence similarity of 94.9 %. Phylogenetic analysis showed that strain NIO-S10T was clustered with members of the genus Tumebacillus and together with members of the genus Effusibacillus. Based on phenotypic characteristics and phylogenetic inference, strain NIO-S10T represents a novel species of the genus Tumebacillus, for which the name Tumebacillus lipolyticus sp. nov. is proposed. The type strain is NIO-S10T ( = KCTC 32289T = MTCC 12483T).

Lunatimonas lonarensis gen. nov., sp. nov., a haloalkaline bacterium of the family Cyclobacteriaceae with nitrate reducing activity.

Novel pinkish-orange pigmented, Gram-negative staining, half-moon shaped, non-motile, strictly aerobic strains designated AK24(T) and AK26 were isolated from water and sediment samples of Lonar Lake, Buldhana district, Maharahstra, India. Both strains were positive for oxidase, catalase and ß-galactosidase activities. The predominant fatty acids were iso-C15:0 (41.5%), anteiso-C15:0 (9.7%), iso-C17:0 3OH (9.6%), iso-C17:1 ω9c (10.2%) and C16:1 ω7c/C16:1 ω6c/iso-C15:0 2OH (summed feature 3) (14.4%). The strains contained MK-7 as the major respiratory quinone, and phosphatidylethanolamine and five unidentified lipids as the polar lipids. Blast analysis of the 16S rRNA gene sequence of strain AK24(T) showed that it was closely related to Aquiflexum balticum, with a pair-wise sequence similarity of 91.6%, as well as to Fontibacter ferrireducens, Belliella baltica and Indibacter alkaliphilus (91.3, 91.2 and 91.2% pair-wise sequence similarity, respectively), but it only had between 88.6 and 91.0% pair-wise sequence similarity to the rest of the family members. The MALDI-TOF assay reported no significant similarities for AK24(T) and AK26, since they potentially represented a new species. A MALDI MSP dendrogram showed close similarity between the two strains, but they maintained a distance from their phylogenetic neighbors. The genome of AK24(T) showed the presence of heavy metal tolerance genes, including the genes providing resistance to arsenic, cadmium, cobalt and zinc. A cluster of heat shock resistance genes was also found in the genome. Two lantibiotic producing genes, LanR and LasB, were also found in the genome of AK24(T). Strains AK24(T) and AK26 were very closely related to each other with 99.5% pair-wise sequence similarity. Phylogenetic analysis indicated that the strains were members of the family Cyclobacteriaceae and they clustered with the genus Mariniradius, as well as with the genera Aquiflexum, Cecembia, Fontibacter, Indibacter, and Shivajiella. DNA-DNA hybridization between strains AK24(T) and AK26 showed a relatedness of 82% and their rep-PCR banding patterns were very similar. Based on data from the current polyphasic study, it is proposed that the isolates be placed in a new genus and species with the name Lunatimonas lonarensis gen. nov., sp. nov. The type strain of Lunatimonas lonarensis is AK24(T) (=JCM 18822(T)=MTCC 11627(T)).

Flavobacterium nitratireducens sp. nov., an amylolytic bacterium of the family Flavobacteriaceae isolated from coastal surface seawater.

A novel Gram-negative, rod-shaped, non-motile bacterium, designated strain N1(T), was isolated from a marine water sample collected from the sea shore, Bay of Bengal, Visakhapatnam, India. The strain was positive for starch hydrolysis, nitrate reduction and ornithine decarboxylase activities and negative for citrate utilization, urease, oxidase, catalase and DNase activities. The predominant fatty acids were C16 : 0 3-OH, iso-C15 : 0, iso-C15 : 0 3-OH, iso-C17 : 0 3-OH, anteiso-C15 : 0, C16 : 0, C15 : 0 3-OH, and C16 : 1ω7c and/or iso-C15 : 0 2-OH (summed feature 3). Strain N1(T) contained menaquinone 6 (MK-6) as the sole respiratory quinone. The only polyamine was homospermidine and the major polar lipids were phosphatidylethanolamine (PE), three unidentified aminolipids (AL1-AL3) and two unidentified lipids (L1, L2). The DNA G+C content of the strain was 36.3 mol%. 16S rRNA gene sequence analysis indicated that strain N1(T) was a member of the genus Flavobacterium and closely related to Flavobacterium resistens with pairwise sequence similarity of 96.5 %. Phylogenetic analysis showed that strain N1(T) clustered with Flavobacterium glycines and Flavobacterium daejeonense with a distance of 4.8 and 6.0 % (95.2 and 94.0 % similarity), respectively. Based on the phenotypic characteristics and on phylogenetic inference, strain N1(T) represents a novel species of the genus Flavobacterium, for which the name Flavobacterium nitratireducens sp. nov. is proposed. The type strain is N1(T) ( = MTCC 11155(T) = JCM 17678(T)).

Photobacterium marinum sp. nov., a marine bacterium isolated from a sediment sample from Palk Bay, India.

The novel, cream colored, Gram-staining-negative, rod-shaped, motile bacteria, designated strains AK15(T) and AK18, were isolated from sediment samples collected from Palk Bay, India. Both strains were positive for arginine dihydrolase, lysine decarboxylase, oxidase, nitrate reduction and methyl red test. The major fatty acids were C16:0, C18:1 ω7c, C16:1 ω7c and/or C16:1 ω6c and/or iso-C15:0 2-OH (summed feature 3). Polar lipids content of strains AK15(T) and AK18 were found to bephosphatidylethanolamine (PE), two unidentified phospholipids (PL1 and PL2) and three unidentified lipids (L1-L3). The 16S rRNA gene sequence analysis indicated strains AK15(T) and AK18 as the members of the genus Photobacterium and closely related to the type strain Photobacterium jeanii with pair-wise sequence similarity of 96.7%. DNA-DNA hybridization between strain AK15(T) and AK18 showed a relatedness of 87%. Based on data from the current polyphasic study, strains AK15(T) and AK18 are proposed as novel species of the genus Photobacterium, for which the name Photobacterium marinum sp. nov. is proposed. The type strain of Photobacterium marinum is AK15(T) (=MTCC 11066(T)=DSM 25368(T)).

Algoriphagus shivajiensis sp. nov., isolated from Cochin back water, India.

Novel orange-pigmented, Gram-negative, rod-shaped, non-motile bacteria, designated strains NIO-S3(T) and NIO-S4, were isolated from a water sample collected from Cochin back waters, Thanneermukkom and Arookutty, Kerala, India. Both strains were positive for oxidase and catalase activities, and hydrolyzed gelatin and Tween 40. The predominant fatty acids were iso-C15:0, anteiso-C15:0, iso-C17:0 3OH, C16:1ω7c/C16:1ω6c (summed feature 3) and iso-C17:1ω9c/C16:0 10-methyl (summed feature 9), whereas MK-7 was the major respiratory quinone, and phosphatidylethanolamine, two unidentified phospholipids and one unidentified lipid were the only polar lipids. The DNA G+C content of the two strains was 43.7 and 43.6mol%, respectively. The 16S rRNA gene sequence analysis indicated that they were members of the genus Algoriphagus and closely related to Algoriphagus olei CC-Hsuan-617(T), Algoriphagus aquatilis A8-7(T), Algoriphagus aquaeductus LMG 24398(T) and Algoriphagus mannitolivorans DSM 15301(T), with pairwise sequence similarities of 96.8, 96.6, 96.2 and 96.2%, respectively. DNA-DNA hybridization between strains NIO-S3(T) and NIO-S4 showed a relatedness of 89%. Based on data from the current polyphasic study, the strains are proposed as a novel species of the genus Algoriphagus, for which the name Algoriphagus shivajiensis sp. nov. is proposed. The type strain of A. shivajiensis is NIO-S3(T) (=JCM 17885(T)=MTCC 11066(T)).

Oceanospirillum nioense sp. nov., a marine bacterium isolated from sediment sample of Palk bay, India.

A novel Gram-negative, spiral shaped, motile bacterium, designated strain NIO-S6(T), was isolated from a sediment sample collected from Off-shore Rameswaram, Tamilnadu, India. Strain NIO-S6(T) was found to be positive for oxidase, DNase and lysine decarboxylase activities and negative for catalase, gelatinase, lipase, ornithine decarboxylase, nitrate reductase, aesculinase, amylase and urease activities. The fatty acids were determined to be dominated by C10:0 3OH, C16:0, C16:1 and C18:1. Strain NIO-S6(T) contains Q-8 as the major respiratory quinone. The DNA G+C content of the strain NIO-S6(T) was determined to be 49.5 ± 0.6 mol %. Phylogenetic analysis based on 16S rRNA gene sequence of strain NIO-S6(T) indicated Oceanospirillum linum and Oceanospirillum maris of the family Oceanospirillaceae (phylum Proteobacteria) are the closest related species with sequence similarities of 98.4 and 97.8 % respectively. Other members of the family showed sequence similarities <96.4 %. However, DNA-DNA hybridization with Oceanospirillum linum LMG 5214(T) and Oceanospirillum maris LMG 5213(T) showed a relatedness of 31.5 and 46.9 % with respect to strain NIO-S6(T). Based on the phenotypic characteristics and on phylogenetic inference, strain NIO-S6(T) is proposed as a novel species of the genus Oceanospirillum as Oceanospirillum nioense sp. nov. and the type strain is NIO-S6(T) (=MTCC 11154(T) = KCTC 32008(T)).

Mariniradius saccharolyticus gen. nov., sp. nov., a member of the family Cyclobacteriaceae isolated from marine aquaculture pond water, and emended descriptions of the genus Aquiflexum and Aquiflexum balticum.

A novel marine, Gram-stain-negative, oxidase- and catalase- positive, rod-shaped bacterium, designated strain AK6(T), was isolated from marine aquaculture pond water collected in Andhra Pradesh, India. The fatty acids were dominated by iso-C15:0, iso-C17:1ω9c, iso-C15:1 G, iso-C17:0 3-OH and anteiso-C15:0. Strain AK6(T) contained MK-7 as the sole respiratory quinone and phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid and seven unidentified lipids as polar lipids. The DNA G+C content of strain AK6(T) was 45.6 mol%. Phylogenetic analysis showed that strain AK6(T) formed a distinct branch within the family Cyclobacteriaceae and clustered with Aquiflexum balticum DSM 16537(T) and other members of the family Cyclobacteriaceae. 16S rRNA gene sequence analysis confirmed that Aquiflexum balticum DSM 16537(T) was the nearest neighbour, with pairwise sequence similarity of 90.1%, while sequence similarity with the other members of the family was <88.5%. Based on differentiating phenotypic characteristics and phylogenetic inference, strain AK6(T) is proposed as a representative of a new genus and species of the family Cyclobacteriaceae, as Mariniradius saccharolyticus gen. nov., sp. nov. The type strain of Mariniradius saccharolyticus is AK6(T) (=MTCC 11279(T)=JCM 17389(T)). Emended descriptions of the genus Aquiflexum and Aquiflexum balticum are also proposed.

Shivajiella indica gen. nov., sp. nov., a marine bacterium of the family "Cyclobacteriaceae" with nitrate reducing activity.

Novel orange pigmented, Gram-negative-staining, rod-shaped, non-motile, strictly aerobic strains designated NIO-S1(T) and NIO-S2 were isolated from the water sample of a pond adjacent to the coast and an algal mat from a fish pond, respectively, at Kakinada, India. Both strains were positive for oxidase, catalase and ß-galactosidase activities. The predominant fatty acids in NIO-S1(T) were iso-C(15:0) (39.6%), anteiso-C(15:0) (9.9%), iso-C(17:0) 3OH (10.9%) and C(16:1)ω7c/C(16:1)ω6c (summed feature 3) (5.7%). The strains contained MK-7 as the major respiratory quinine, and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unidentified lipids as the polar lipids. Phylogenetic analysis indicated that strain NIO-S1(T) was a member of the family "Cyclobacteriaceae" of the class "Sphingobacteriia" and it clustered with the genera Fontibacter, Cecembia and Aquiflexum with phylogenetic distances of 6.8, 9.0 and 12.2% (93.2, 91.0 and 87.8% similarity), respectively. DNA-DNA hybridization between strains NIO-S1(T) and NIO-S2 showed a relatedness of 93% and rep-PCR banding patterns were similar. Based on data from the current polyphasic study, it is proposed that the new isolates be placed in a new genus and species with the name Shivajiella indica gen. nov., sp. nov. The type strain of Shivajiella indica is NIO-S1(T) (= KCTC 19812(T)=MTCC 11065(T)).