FROM THE PERSPECTIVE OF THE TARGET A CIS-REGULATORY PERSPECTIVE ON THE EFFECTS OF MICROGRAVITY PROJECT SUMMARY: LONG DURATION SPACEFLIGHT AND FUTURE COLONIZATION MISSIONS WILL REQUIRE PLANTS TO PROVIDE MUCH NEEDED RESOURCES FOR HUMAN HEALTH AND WELL BEING. PLANTS ARE AMAZINGLY HARDY ORGANISMS WHICH HAVE OVER THE YEARS DEVELOPED MECHANISMS TO PERCEIVE AND RESPOND TO STRESSES THEY ENCOUNTER. THIS EVOLUTION OCCURRED IN TERRESTRIAL ENVIRONMENTS HERE ON EARTH AND NOT IN THE ENVIRONMENT OF SPACE. WE ARE INTERESTED IN UNDERSTANDING THE MECHANISMS BY WHICH PLANTS PERCEIVE AND RESPOND TO STRESSES SUCH AS A CHANGE IN GRAVITY OR OTHER ASPECTS OF SPACE FLIGHT. UNDERSTANDING HOW PLANTS SENSE AND RESPOND TO THIS ENVIRONMENT IS CRITICAL FOR ENABLING PLANT GROWTH IN SPACE. WE EXAMINED THE AVAILABLE TRANSCRIPTIONAL DATA IN GENE LAB AND OBSERVED THAT WHILE THERE WERE FEW GENES THAT WERE COHESIVELY EXPRESSED ACROSS ALL CONDITIONS THERE WAS SOME CONSISTENCY IN THE CIS-REGULATORY FEATURES THAT ARE ENRICHED. BASED ON FINDINGS FROM GENELAB TRANSCRIPTIONAL EXPERIMENTS WE PROPOSE TO EXAMINE THE EFFECTS OF SPACE FLIGHT THROUGH THE LENS OF THE CIS-REGULATORY NETWORK. OUR ANALYSIS OF THE ARABIDOPSIS TRANSCRIPTIONAL DATASETS AVAILABLE IN GENELAB SHOWS THAT THERE IS LITTLE OVERLAP IN THE INDIVIDUAL DIFFERENTIALLY EXPRESSED (DE) GENES BETWEEN THE VARIOUS SPACEFLIGHT EXPERIMENTS. RATHER THAN QUERYING THE OVERLAP OF SPECIFIC GENES ACROSS EXPERIMENTS WE ARE FOCUSING ON UPSTREAM SIGNALING COMPONENTS USING THE CONSERVED CIS-REGULATORY SEQUENCES AS A TOOL TO UNCOVER THE UPSTREAM REGULATORS. WE HAVE IDENTIFIED ENRICHED AND CONSERVED MOTIFS IN THE PROMOTERS OF GENES DIFFERENTIALLY EXPRESSED IN SPACEFLIGHT EXPERIMENTS THIS APPROACH IS INCLUSIVE AND PARTICULARLY BENEFICIAL FOR IDENTIFYING PATTERNS IN LIMITED AND HIGHLY DISSIMILAR DATA SETS. WE WILL TAKE A FOCUSED APPROACH SELECTING A FEW OF THE MOTIFS THAT ARE ENRICHED IN GENES THAT ARE RESPONSIVE TO MICROGRAVITY ACROSS MULTIPLE INDEPENDENT EXPERIMENTS IN GENELAB AND EXAMINE THE CIS-REGULATORY CONTROL FROM THE BOTTOM UP. THE OBJECTIVE OF THIS STUDY IS TO UTILIZE THE GENELAB REPOSITORY AS A STARTING POINT TO IDENTIFY MECHANISMS REGULATING PLANT RESPONSES TO SPACEFLIGHT AND MICROGRAVITY. OUR SPECIFIC AIMS ARE: 1. TO SCREEN A Y1H TRANSCRIPTION FACTOR LIBRARY TO IDENTIFY SPECIFIC TRANSCRIPTION FACTORS 2. CONFIRM DNA-BINDING OF TRANSCRIPTION FACTORS AND OBTAIN TRANSCRIPTION FACTOR MUTANTS 3. GENERATE PROMOTER:REPORTER LINES TO TEST IN PLANTA THIS PROPOSAL IS IN RESPONSE TO APPENDIX G OF THE RESEARCH OPPORTUNITIES IN SPACE BIOLOGY RESEARCH ANNOUNCEMENT. SPECIFICALLY OUR PROPOSAL WILL ADDRESS 4B) SYSTEMS BIOLOGY EXPERIMENTAL RESEARCH USING THE GENELAB DATA SYSTEM UNDER "INVESTIGATORS WHO WISH TO DEVELOP NEW EXPERIMENTAL HYPOTHESES BASED ON DATA IN THE GENELAB DATABASE". WE ANTICIPATE THAT THIS WORK WILL ENABLE US TO DELINEATE SIGNALING PATHWAY/S RESPONDING TO MICROGRAVITY AND YIELD NEW KNOWLEDGE ON PLANT SPECIFIC RESPONSES TO SPACEFLIGHT.