MS1. Title Page: should include the title, name of each author, and the

institution at which the work was performed. • Text should be centered.

DWA3, an Arabidopsis DWD protein, acts as a negative regulator in ABA signal

transduction   Jae-Hoon Lee, a William Terzaghi,a,b and Xing Wang Deng a,*

 

a Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520-8104, U.S.A.

b Department of Biology, Wilkes University, Wilkes-Barre, PA 18766, U.S.A.

  

* Corresponding author, Fax: +1 203 432 3204 E-mail address: xingwang.deng@yale.edu Keywords: CUL4-based E3 ligase; DWD; ABA; negative regulator;

signal transduction pathway; Arabidopsis

MS2. Abstract: summarizes the paper in 250 words or less.

DWD proteins have been reported as substrate receptors for cullin–RING ubiquitin ligase 4 (CRL4). Upon screening T-DNA mutants of DWD genes for abscisic acid (ABA) responses we obtained several candidates which exhibited ABA-hypersensitivity and one was named DWA3 (DWD hypersensitive to ABA 3). DWA3 associated with the CRL4 components DDB1 and CUL4, indicating that DWA3 may function as a substrate receptor for CRL4. ABA-inducible transcription factors (ABI5 and AtMYC2) and their downstream genes were hyper-induced by ABA in dwa3. Taken together, we suggest DWA3 is a negative regulator of ABA responses and may be involved in protein degradation mediated by CRL4.

MS2. Abstract: summarizes the paper in 250 words or less.• mini-papers that are often published separately

DWD proteins have been reported as substrate receptors for cullin–RING ubiquitin ligase 4 (CRL4). Upon screening T-DNA mutants of DWD genes for abscisic acid (ABA) responses we obtained several candidates which exhibited ABA-hypersensitivity and one was named DWA3 (DWD hypersensitive to ABA 3). DWA3 associated with the CRL4 components DDB1 and CUL4, indicating that DWA3 may function as a substrate receptor for CRL4. ABA-inducible transcription factors (ABI5 and AtMYC2) and their downstream genes were hyper-induced by ABA in dwa3. Taken together, we suggest DWA3 is a negative regulator of ABA responses and may be involved in protein degradation mediated by CRL4.

MS2. Abstract: summarizes the paper in 250 words or less. •mini-papers that are often published separately •should have 1 or 2 sentences of Introduction

DWD proteins have been reported as substrate receptors for cullin–RING ubiquitin ligase 4 (CRL4). Upon screening T-DNA mutants of DWD genes for abscisic acid (ABA) responses we obtained several candidates which exhibited ABA-hypersensitivity and one was named DWA3 (DWD hypersensitive to ABA 3). DWA3 associated with the CRL4 components DDB1 and CUL4, indicating that DWA3 may function as a substrate receptor for CRL4. ABA-inducible transcription factors (ABI5 and AtMYC2) and their downstream genes were hyper-induced by ABA in dwa3. Taken together, we suggest DWA3 is a negative regulator of ABA responses and may be involved in protein degradation mediated by CRL4.

MS2. Abstract: summarizes the paper in 250 words or less. •mini-papers that are often published separately •should have 1 or 2 sentences of Introduction•1- 2 sentences of Materials and MethodsDWD proteins have been reported as substrate receptors for cullin–RING ubiquitin ligase 4 (CRL4). Upon screening T-DNA mutants of DWD genes for abscisic acid (ABA) responses we obtained several candidates which exhibited ABA-hypersensitivity and one was named DWA3 (DWD hypersensitive to ABA 3). DWA3 associated with the CRL4 components DDB1 and CUL4, indicating that DWA3 may function as a substrate receptor for CRL4. ABA-inducible transcription factors (ABI5 and AtMYC2) and their downstream genes were hyper-induced by ABA in dwa3. Taken together, we suggest DWA3 is a negative regulator of ABA responses and may be involved in protein degradation mediated by CRL4.

MS2. Abstract: summarizes the paper in 250 words or less. •mini-papers that are often published separately •should have 1 or 2 sentences of Introduction•1- 2 sentences of Materials and Methods•3-4 sentences of Results, including quantitative data

MS2. Abstract: summarizes the paper in 250 words or less. •mini-papers that are often published separately •should have 1 or 2 sentences of Introduction•1- 2 sentences of Materials and Methods•3-4 sentences of Results, including quantitative data •1-2 sentences of Discussion

MS2. Abstract: summarizes the paper in 250 words or less. •mini-papers that are often published separately •should have 1 or 2 sentences of Introduction•1- 2 sentences of Materials and Methods•3-4 sentences of Results, including quantitative data •1-2 sentences of Discussion•rarely cite references. If they do, full citation must be included.

MS3. Introduction: explains why the experiment was done.

MS3. Introduction: explains why the experiment was done. •provides enough detail about what was previously known to explain what the outstanding questions are

MS3. Introduction: explains why the experiment was done. •provides enough detail about what was previously known to explain what the outstanding questions are•specifically states the hypothesis that was tested.

MS3. Introduction: explains why the experiment was done. •provides enough detail about what was previously known to explain what the outstanding questions are•specifically states the hypothesis that was tested.•I write it last, to guide readers to my results

MS4. Materials and Methods: provide sufficient detail that another biologist could repeat your experiment.

MS4. Materials and Methods: provide sufficient detail that another biologist could repeat your experiment. •list the organisms used (including the latin binomial) the names of the reagents, procedures followed, etc

MS4. Materials and Methods: provide sufficient detail that another biologist could repeat your experiment. •list the organisms used (including the latin binomial) the names of the reagents, procedures followed, etc•do not give the recipe for each reagent.

•Instead, cite the reference from which the recipe was obtained.

MS4. Materials and Methods: provide sufficient detail that another biologist could repeat your experiment. •list the organisms used (including the latin binomial) the names of the reagents, procedures followed, etc•do not give the recipe for each reagent. •Do cite the manufacturers of esoteric reagents or equipment

MS4. Materials and Methods: provide sufficient detail that another biologist could repeat your experiment. •list the organisms used (including the latin binomial) the names of the reagents, procedures followed, etc•do not give the recipe for each reagent. •Do cite the manufacturers of esoteric reagents or equipment•Both Materials and Methods and Results should be written in the past tense. Routine calculations are not described here, unless they are done in an unusual way.

MS5. Results: devote one paragraph to each figure or table

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods•third sentence states where the results are presented

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods•third sentence states where the results are presented•remaining sentences point out the key features.

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods•third sentence states where the results are presented•remaining sentences point out the key features. •Results can be presented as figures or tables, which should be presented on separate pages attached after the “literature cited”

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods•third sentence states where the results are presented•remaining sentences point out the key features. •Results can be presented as figures or tables, which should be presented on separate pages attached after the “literature cited”•each figure or table should have its own title and caption

MS5. Results: devote one paragraph to each figure or table•start with a sentence explaining the purpose of the expt•second sentence should summarize the methods•third sentence states where the results are presented•remaining sentences point out the key features. •Results can be presented as figures or tables, which should be presented on separate pages attached after the “literature cited”•each figure or table should have its own title and caption•caption gives sufficient information that a reader can figure out what it is about without reading the text. i.e. summarizes M&M & identifies panels, symbols, etc

MS6. Discussion: one paragraph per figure or table + a global discussion at end

MS6. Discussion: one paragraph per figure or table + a global discussion at end•First sentence summarizes results

MS6. Discussion: one paragraph per figure or table + a global discussion at end•First sentence summarizes results•Remaining sentences explain them, and may propose ways to test these explanations

MS6. Discussion: one paragraph per figure or table + a global discussion at end•First sentence summarizes results•Remaining sentences explain them, and may propose ways to test these explanations•Last paragraph discusses global implications: now that we know this, how does it change our world?

MS7. Literature cited: all citations listed in the text should be listed at the end of the paper.

MS7. Literature cited: all citations listed in the text should be listed at the end of the paper. •Formats for citing and listing references vary among journals.

MS7. Literature cited: all citations listed in the text should be listed at the end of the paper. •Formats for citing and listing references vary among journals. •Use the format (Smith, 2008) in the text

MS7. Literature cited: all citations listed in the text should be listed at the end of the paper. •Formats for citing and listing references vary among journals. •Use the format (Smith, 2008) in the text •Use this format in the “Literature Cited:” Smith, E.J. (2008) BRF3 encodes a novel ubiquitin ligase. Molecular Plant 3: 345-361

Manuscript Draft Grading Checklist1) Abstract (6 pts)

Were all elements (I, M&M, R & D) present?Were all elements adequately explained?Was all information relevant?Was all information clearly and succinctly explained?

2) Introduction (6 pts)Was the hypothesis (or purpose) clearly stated?Was adequate background information provided?Was all information relevant?Was all information clearly explained?

3) Materials and Methods (4 pts)Were all procedures clearly and accurately explained?Was all information relevant?

4) Results (10 pts)Was there a separate paragraph for each expt? Did the first sentence of each para state the purpose?Did the second sentence summarize the methods?Did the third sentence state where the results are? Did the remaining sentences explain the figure/table and point

out key results? Did figures and tables do the job?Titles and captions?

5) Discussion (8 pts)Were key results summarized?Were key results discussed?Were further experiments proposed?Were broader implications discussed?

6) Literature cited (2 pts)Were references used correctly in the text?Were all citations made in text listed in correct format?Were any references not cited in the text?

7) Writing (4 pts)OrganizationClarityConcisenessGrammar and spelling