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The Arabidopsis ClpB/Hsp100 family of proteins: chaperones for stress and chloroplast development

Lee, Ung,Rioflorido, Ignatius,Hong, Suk-Whan,Larkindale, Jane,Waters, Elizabeth R.,Vierling, Elizabeth Blackwell Publishing Ltd 2007 The Plant journal Vol.49 No.1
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Summary The Casein lytic proteinase/heat shock protein 100 (Clp/Hsp100) proteins are chaperones that act to remodel/disassemble protein complexes and/or aggregates using the energy of ATP. In plants, one of the best-studied proteins from this family is cytosolic ClpB1 (At1g74310), better known in Arabidopsis as AtHsp101, which is a heat shock protein required for acclimation to high temperatures. Three other ClpB homologues have been identified in the Arabidopsis genome (ClpB2, ClpB3 and ClpB4; At4g14670, At5g15450 and At2g25140). To define further the roles of these chaperones in plants we investigated their intracellular localization, evolutionary relationships, patterns of expression and the phenotypes of corresponding T-DNA insertion mutants. We first found that ClpB2 was misannotated; there is no functional ClpB/Hsp100 gene at this locus. By fusing the putative transit peptides of ClpB3 and ClpB4 with GFP, we showed that these proteins are targeted to the chloroplast and mitochondrion, respectively, and we therefore designated them as ClpB-p and ClpB-m. Phylogenetic analysis supports two major lineages of ClpB proteins in plants, an ‘eukaryotic’, cytosol/nuclear-localized group containing AtHsp101, and an organelle-localized lineage, containing both ClpB-p and ClpB-m. Although AtHsp101, ClpB-p and ClpB-m transcripts all accumulate dramatically at high temperatures, the T-DNA insertion mutants of ClpB-p and ClpB-m show no evidence of seedling heat stress phenotypes similar to those observed in AtHsp101 mutants. Strikingly, ClpB-p knockouts were seedling lethals, failing to accumulate chlorophyll or properly develop chloroplasts. Thus, in plants, the function of ClpB/Hsp100 proteins is not restricted to heat stress, but a specific member of the family provides housekeeping functions that are essential to chloroplast development.
2
Identification of ROS1 rearrangement in gastric adenocarcinoma

Lee, Jeeyun,Lee, Seung Eun,Kang, So Young,Do, In‐,Gu,Lee, Sujin,Ha, Sang Yun,Cho, Jeonghee,Kang, Won Ki,Jang, Jiryeon,Ou, Sai‐,Hong Ignatius,Kim, Kyoung‐,Mee Wiley Subscription Services, Inc., A Wiley Company 2013 Cancer Vol.119 No.9
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AbstractBACKGROUND:Recently, chromosomal rearrangements involving receptor tyrosine kinases (RTKs) have been described in common epithelial malignancies, including nonsmall cell lung cancer (NSCLC), colorectal cancer, and breast cancer. One of these RTKs, c‐ros oncogene 1, receptor tyrosine kinase (ROS1), has been identified as a driver mutation in NSCLC, because its inhibition by crizotinib, an anaplastic lymphoma receptor tyrosine kinase (ALK)/met proto‐oncogene hepatocyte growth factor receptor (MET)/ROS1 inhibitor, led to significant tumor shrinkage in ROS1‐rearranged NSCLC. Currently, only human epidermal growth factor 2 (HER2)‐targeted therapy in combination with chemotherapy has been successful in significantly prolonging the survival of patients with advanced gastric cancer (GC). There is a need for the discovery of additional novel targets in GC.METHODS:Anti‐ROS1 immunohistochemistry (IHC) was used to screen 495 GC samples and was followed by simultaneous ROS1 break‐apart fluorescence in situ hybridization (FISH) and reverse transcriptase‐polymerase chain reaction (RT‐PCR) analyses in IHC‐positive samples. Fusion partners in ROS1‐rearranged GC were determined by RT‐PCR. In all 495 samples, HER2 amplification was identified with FISH, and MET expression was identified by IHC.RESULTS:Twenty‐three tumor samples were ROS1 IHC‐positive. Three of 23 patients were ROS1 FISH positive, HER2 FISH negative, and negative for MET overexpression; and 2 of those 3 patients harbored a solute carrier family 34 (sodium phosphate), member 2 (SLC34A2)‐ROS1 fusion transcripts. No fusion partner was identified in the third patient. Both patients who had SLC34A2‐ROS1 transcripts had poorly differentiated histology with recurrence and death within 2 years of curative surgery. ROS1 IHC‐positive status was not identified as an independent prognostic factor for overall survival.CONCLUSIONS:In this study, an SLC34A2‐ROS1 rearrangement was identified in GC, and the results provide a rationale for investigating the clinical efficacy of ROS1 inhibitors in this unique molecular subset of GC. Cancer 2013. © 2013 American Cancer Society.

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