Citation
Poster Sessions: Biological Sciences

Material Information

Title:
Poster Sessions: Biological Sciences
Abbreviated Title:
Quest
Creator:
Buckler, Sarah
Fingar, Emily
Flores, Tiffany
Goodman, Brooke
Mateo, Nurys
Roberts, Kelsey
Salerno, Robert
Salinas, Mariabelen
Sard, Nicholas
Publication Date:
Copyright Date:
2021

Notes

Abstract:
Comparing Song Repertoires in Urban vs. Rural Northern Cardinals by Brooke Goodman. ( ,,,,,,,, )
Abstract:
Late Pleistocene Frog Fossils from Cathedral Cave, Nevada by Mariabelen Salinas.
Abstract:
Genetic Screening to Find Novel Regulators of Tumor Suppressor Homolog Kinase Responsive to Stress B (KrsB) by Emily Fingar.
Abstract:
Investigation of The Role of KrsB in Rap1-Mediated Adhesion of Dictyostelium discoideum by Kelsey Roberts.
Abstract:
Regulation of Ras-Associated Protein-1 By Kinase Responsive to Stress B in Dictyostelium discoideum by Tiffany Flores.
Abstract:
Mechanism of filamin action in response to mechanical stimuli by Sarah Buckler.
Abstract:
A Morphometrics Study of Prostrate Leaves of Wintergreen Fern Dryopteris intermedia by Robert Salerno.
Abstract:
Skull histology of anole lizards by Nurys Mateo.
Summary:
Session Chair: Nicholas Sard
Acquisition:
Collected for SUNY Oswego Institutional Repository by the online self-submittal tool. Submitted by Zach Vickery.

Record Information

Source Institution:
SUNY Oswego Institutional Repository
Holding Location:
SUNY Oswego
Rights Management:
All rights reserved by the source institution.

OswegoDL Membership

Aggregations:
Quest
Added automatically

Downloads

This item is only available as the following downloads:


Full Text

PAGE 1

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



PAGE 1

Late Pleistocene Frogs From Cathedral Cave, Nevada Maria Belen Salinas Department of Biological Sciences, SUNY Oswego Abstract Pleistocene microvertebrae fossils of frogs and toads are reported from Cathedral Cave (CC), White Pine County, Nevada. Skeletal remains include 5 ilia belonging to at least 3 species of amphibians, including one extinct species; all occur in or ad jacent to the area today. These provide valuable information about the taxonomic diversity and paleoecology of the Late Pleistocene fauna of east central Nevada. Background In 1989, CC was excavated to determine its paleoecological significance [4]. The site is located in White Pine County, Nevada, in the Great Basin of the western United States. The locality has a maximum age range of 146.02 ± 2.584 ka to 151.2 ± 4.4 ka [3]. Identification of amphibian specimens from CC have not received the same attention given to mammals [4]. Because of the need for more investigations into different groups, I examined anuran ilia excavated from CC. Due to intraspecific variation in Anura, the ilium, a pelvic bone, has become valuable for identification [6]. Methods Anuran specimens were previously sorted and identified preliminarily. My goal was to further refine those identifications. Specimens were examined under a dissecting microscope. Images were taken with a Canon Rebel XSi camera attached to a microscope. Figures were designed using GIMP 2.10.22. Conclusions The deposit provides important information for understanding the composition of the region today as a product of previous biotic factors [4]. Of extant species identified, Anaxyrus punctatus, cognatus, boreas , and speciosus inhabit the southwestern United States. The extinct, A. pliocompactilis , also was recovered from the WaKeeny Local Fauna, Trego County, Kansas, [2]. Despite no records of A. pliocompactilis in the Pleistocene, [5] suggests the species has inhabited areas similar to A. punctatus , which is commonly found in Nevada today. These findings provide valuable paleo ecological information that can be used to infer the environmental conditions CC previously experienced. In order to perform accurate identifications, a regional framework containing fossil data from other sites must be created to further compare data [4]. References 1. J. Arthur Thomson, M.A., LL.D. Outlines of Zoology (New York, NY: D. Appleton & Company, 1916) 2. Holman, J. (2003). Fossil Frogs and Toads of North America, Indiana University Press , 117 149. ISBN: 0 253 34280 5 3. Jass , C. N. (2009). Pleistocene Lagomorphs from Cathedral Cave, Nevada. PaleoBios , 29 (1), 1 12. 4. Jass , C. N., & C. J. Bell. (2011) Arvicoline Rodent Fauna from the Room 2 Excavation in Cathedral Cave, White Pine County, Nevada, and its Biochronologic Significance, Journal of Vertebrate Paleontology, 31 (3), 684 699. 5. Parmley , D., Chandler, R., & Chandler, L. (2015). Fossil Frogs of the Late Clarendonian (Late Miocene) Pratt Slide Local Fauna of Nebraska, with the Description of a New Genus. Journal of Herpetology, 49 (1), 143 149. 6. Gómez, R., & Turazzini , G. (2016). An Overview of the Ilium of Anurans (Lissamphibia, Salientia), with a Critical Appraisal of the Terminology and Pri mary Homology of Main Ilial Features. Journal of Vertebrate Paleontology, 36 (1), 1 12. Figure 2. Ilia in lateral view; scale bar = 1 mm. A , left ilium; Anaxyrus pliocompactilis , cognatus, or speciosus (?) (specimen 153); B D , right ilia; A. punctatus (151, 154, 155, respectively); E , right ilium; A. boreas (?) (152). Abbreviations: acf , acetabular fossa; acr , acetabular rim; dae , dorsal acetabular expansion; dpm , dorsal prominence; dpt , dorsal protuberance; ish , ilial shaft; vae , ventral acetabular expansion. Skeleton drawing from [1]. Figure 1. Geographic Location of CC [4] Acknowledgements I would like to thank Dr. C. Sagebiel at the University of Texas at Austin for providing specimens and Drs. C. Jass , C. Bell, and J. Mead for providing preliminary identifications and feedback on images.



PAGE 1

Mechanism of Filamin Action in Response to Mechanical Stimuli Sarah Buckler, Yulia Artemenko Department of Biological Sciences, SUNY Oswego, Oswego NY J Dictyostelium cells (WT, filamin null) expressing mCherry tagged filamin and GFP tagged Ras binding domain (RBD) Image with epifluorescence every 3 sec for 20 frames After 5 frames, stimulate cells with shear flow for 2 sec at 50 mbar pressure Ibidi µ Slide III 3in1 INTRODUCTION Dictyostelium discoideum Social amoeba 1 Contains many genes homologous to higher eukaryotes 1 Useful for studying cell motility, chemotaxis, signal transduction, etc. 1 Directed Cell Migration Cells respond to various stimuli, including chemical and mechanical cues. 2 Activation of the signal transduction network can bias actin polymerization allowing for directed cell migration. 3 Although chemical and mechanical stimuli appear to activate similar signal transduction networks, how cells sense mechanical stimuli remains unclear . 3 An intact actin cytoskeleton is necessary for cell response to mechanical stimuli. 3 Actin binding Protein Filamin Crosslink actin filaments 2 Stabilize 3D actin webs 5 Link actin to plasma membrane 5 Implicated in sensing mechanical pressure 4 Previously shown in our lab that response to acute mechanical stimulation is reduced in the absence of filamin in D.discoideum CONCLUSIONS AND FUTURE DIRECTIONS REFERENCES stimuli. Filamin is not involved in the response to chemical stimuli. Effects of filamin are not due to altered adhesion of filamin null cells to the surface (data not shown) . 1. dictyBase . Dictyostelium discoideum : Model System in Motion. http://dictybase.org/tutorial/ (accessed Aug 18, 2020). 2. Pollard, T. D. (2016). Actin and Actin Binding Proteins. Cold Spring Harbor Perspectives in Biology, 8 (8). doi:10.1101/cshperspect.a018226 3. Artemenko , Y., Devreotes , P. N. Assessment of Dictyostelium discoideum Response to Acute Mechanical Stimulation. J. Vis. Exp. (129), e56411, doi:10.3791/56411 (2017). 4. Luo, T., Mohan, K., Iglesias, P. et al. Molecular mechanisms of cellular mechanosensing . Nature Mater 12, 1064 1071 (2013). https://doi.org/10.1038/nmat3772 5. Sunderland, M. E., " Dictyostelium discoideum ". Embryo Project Encyclopedia (2009 06 10). ISSN: 1940 5030 ACKNOWLEDGMENTS This work was supported by NSF RUI grant no. 1817378 (to Y.A.) and RISE travel funds (to S.B.) Figure 5. Response to stimulation with folic acid is not affected by lack of filamin. Vegetative filamin null cells expressing RBD GFP, as well as empty vector ( pDRH ) or mCherry tagged filamin were stimulated with 100 µM folic acid at time 0. Response was measured as an inverse of drop in cytosolic intensity of RBD GFP. No significant differences were found between the cell lines. RESULTS ABSTRACT Molecular mechanisms by which cells sense and directionally migrate in response to mechanical perturbation, which is critical in homeostasis and many diseases, are not well understood. Dictyostelium discoideum cells exposed to a brief burst of shear flow show rapid and transient activation of multiple components of the signal transduction network, a response that requires an intact actin cytoskeleton of the cell. However, exactly what aspect of the actin cytoskeleton network is responsible for sensing and/or transmitting the signal is unclear. Previous data from our laboratory suggested that actin crosslinking protein filamin is involved in the ability of cells to respond to shear flow. In this study we further characterized the mechanism of filamin action in this response. Filamin itself showed rapid and transient relocalization from the cytosol to the cortex following 2 sec stimulation with shear flow. To detect activation of the signal transduction network in the presence or absence of this actin binding protein, we used fluorescently tagged Ras binding domain biosensor that detects active Ras and was previously shown to relocalize to the cortex following mechanical stimulation. Reduced responsiveness of the network to stimulation with shear flow in the absence of filamin was specific to mechanical stimuli since response to global stimulation with a chemoattractant was comparable between cells with or without filamin. To understand how filamin might be regulating shear flow induced responses we generated truncation constructs of filamin lacking either the actin binding domain or the dimerization domain. Studies are underway to determine whether these truncation constructs are able to rescue the reduced response of filamin null cells to brief stimulation with shear flow, which will offer insight into the molecular mechanism of filamin action in this context. APPROACH binding domain and/or dimerization domain is needed for filamin mediated response to mechanical stimulation in Dictyostelium discoideum . HYPOTHESIS Collect vegetative cells grown on a bacterial lawn, wash in buffer, and plate https://www.mechanobio.info/cytoskeleton dynamics/actin crosslinking/ Figure 2 . Filamin no ABD transiently relocalizes to the cortex in response to acute stimulation with shear flow. Wild type cells expressing mCherry tagged filamin or filamin lacking ABD were imaged with epifluorescence microscopy every 3 sec. Cells were stimulated with shear flow for 2 sec at time 0. (C) Representative images showing increased accumulation of filamin lacking ABD at the cortex 6 sec after stimulation. (D) Response was quantified as the inverse of drop in cytosolic intensity of RFP filamin. Figure 1. Filamin no DD transiently relocalizes to the cortex in response to acute stimulation with shear flow. Wild type cells expressing mCherry tagged filamin or filamin lacking DD were imaged with epifluorescence microscopy every 3 sec. Cells were stimulated with shear flow for 2 sec at time 0. (A) Representative images showing increased accumulation of filamin lacking DD at the cortex 9 sec after stimulation. (B) Response was quantified as the inverse of drop in cytosolic intensity of mCherry filamin signal . Figure 3. Response to acute stimulation with shear flow appears to be reduced in filamin null cells expressing filamin no ABD compared to cells with wild type filamin. Vegetative filamin null cells expressing RBD GFP, as well as empty vector ( pDRH ), mCherry tagged filamin or filamin lacking ABD were stimulated with shear flow for 2 sec at time 0. RBD GFP response ( relocalization from the cytosol to the cortex) was quantified as the inverse of drop in cytosolic intensity. *P<0.05 for vector vs. filamin. binding domain and dimerization stimulation with shear flow. Filamin null cells expressing filamin without ABD or DD responded similarly to cells expressing empty vector, suggesting ABD and DD are required for filamin mediated response of cells to shear flow, although the response was also not significantly different between cells with full length filamin vs. filamin with no ABD. response. binding domain and dimerization domain may not be needed for the re localization to the cortex in response to shear flow stimulation In wild type cells filamin without ABD or DD translocated to the cell cortex similarly to full length filamin in response to shear flow. More data is needed to confirm the role of these domains in Popowicz et al., 2004 . * mean ± SE 0 sec 9 sec 15 sec A A Figure 4. Response to brief stimulation with shear flow is similar in filamin with no DD compared to cells with empty vector. Vegetative filamin null cells expressing RBD GFP, as well as empty vector ( pDRH ), or mCherry tagged filamin lacking DD were stimulated with shear flow for 2 sec at time 0. RBD GFP response ( relocalization from the cytosol to the cortex) was quantified as the inverse of drop in cytosolic intensity. No significant differences were found between the cell lines. mean ± SE 0.8 0.9 1.0 1.1 1.2 1.3 -15 -12 -9 -6 -3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 Filamin Response Time (sec) Filamin (n=11) Filamin-no DD (n=16) B B 0.9 1.0 1.1 1.2 1.3 -15 -12 -9 -6 -3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 RBD GFP Response Time (sec) vector (n=21) Filamin (n=19) Filamin-no ABD (n=18) 0.9 1.0 1.1 1.2 -15 -12 -9 -6 -3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 RBD GFP Response Time (sec) vector (n=18) Filamin-no DD (n=16) mean ± SE mean ± SE mean ± SE mean ± SE * 0.8 0.9 1.0 1.1 1.2 1.3 -15 -12 -9 -6 -3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 Filamin Response Time (sec ) Filamin (n=8) Filamin-no ABD (n=18) mean ± SE 0.9 1.0 1.1 1.2 -15 -12 -9 -6 -3 0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 RBD GFP Response Time (sec) vector (n=29) mCherry-filamin (n=29) 0 sec 6 sec 12 sec mCherry Filamin no DD mCherry Filamin no AB D



PAGE 1

&RPSDULQJ6RQJ5HSHUWRLUHVLQ8UEDQYV5XUDO1RUWKHUQ&DUGLQDOV %URRNH*RRGPDQDQG'DQLHO7%DOGDVVDUUH 'HSDUWPHQWRI%LRORJLFDO6FLHQFHV681<2VZHJR ‡1RUWKHUQ&DUGLQDO &DUGLQDOLVFDUGLQDOLV )LJXUHVRQJLVPDVNHGE\DQWKURSRJHQLF QRLVHDQGXUEDQPDOHVDGMXVWWKHLUVRQJ IUHTXHQFLHVDFFRUGLQJO\6SLHU ‡6RQJIUHTXHQF\DQGOHQJWKLVSRVLWLYHO\ FRUUHODWHGZLWKXUEDQL]DWLRQEXWWKHHIIHFW RIXUEDQL]DWLRQRQUHSHUWRLUHVL]HLVXQNQRZQ1DUDQJR5RGHZDOG ‡&DUGLQDOVVLQJEHWZHHQVRQJVW\SHV6DOHVV,QFUHDVHGVRQJ UHSHUWRLUHVL]HPD\JLYHXUEDQFDUGLQDOVDFRPSHWLWLYHDGYDQWDJHVLQQRLV\ HQYLURQPHQWV ‡ +HUHZHLQWURGXFHDPHWKRGIRUTXDQWLI\LQJFDUGLQDOVRQJUHSHUWRLUH VL]HWRGHWHUPLQHLIXUEDQPDOHVVLQJPRUHVRQJW\SHVLQUHVSRQVHWR DQWKURSRJHQLFQRLVH ,QWURGXFWLRQ )LHOGZRUN ‡-XQH$XJXVWFDSWXUHGDQGEDQGHGELUGVDW%DUU\3DUN6\UDFXVH %3XUEDQ)LJXUHDQG5LFH&UHHN)LHOG6WDWLRQ5&)6UXUDO)LJXUH ‡5HFRUGHGKRXUVRIDXGLRSHUPDOHXVLQJ$XWRPDWHG5HFRUGLQJ8QLWV $58)LJXUH ‡6DPSOHVL]HV%3 PDOHV5&)6 PDOHV ‡$QDYHUDJHRIVRQJVZHUHGHWHFWHGE\WKH$58GDLO\ )LJXUH%DUU\3DUNLQ6\UDFXVH1< )LJXUH5LFH&UHHN)LHOG6WDWLRQLQ2VZHJR1< )LJXUH1RUWKHUQ&DUGLQDO KWWSVXDJRDW*-/ 7ULOO% 7ULOO$ ‡$58UHFRUGLQJVDUHDQDO\]HGXVLQJ.DOHLGRVFRSH3UR6RIWZDUHZKLFKFDQEH WUDLQHGWRUHFRJQL]HDQGFOXVWHUDERXWVRQJW\SHVXVLQJDFODVVLILHU ‡$FODVVLILHULVEXLOWXVLQJH[DPSOHVRIWKHWDUJHWVRQJVDQGVHSDUDWHFODVVLILHUV PXVWEHPDGHIRUWKHGLIIHUHQWILHOGVLWHVGXHWRGLIIHUHQFHVLQVRQJW\SHV ‡6RQJVDUHGLIIHUHQWLDWHGIURPHDFKRWKHUE\WULOODQGV\OODEOHXVDJH ‡6RQJVFDQEHFRPSRVHGRIRQHRUPXOWLSOHWULOOV)LJXUHDQGWULOOVFDQEHUHSHDWHG WKURXJKRXWDELUGVUHSHUWRLUH ‡1RWDOOYRFDOL]DWLRQVDUHVRQJVDQGGHFLGLQJLIDYRFDOL]DWLRQLVDVRQJGHSHQGV XSRQKRZRIWHQLWLVVXQJ )LJXUH$Q$58GHSOR\HGRQDPDOHFDUGLQDO¶V WHUULWRU\ ‡&UHDWHFODVVLILHUVIRUERWK5&)6UXUDODQG%3XUEDQ ‡$QDO\]H$58UHFRUGLQJVRIDQGEXLOGUHSHUWRLUHVIRUDOOLQGLYLGXDOV ‡&RPSDUHVL]HRIVRQJUHSHUWRLUHVRIXUEDQYVUXUDOFDUGLQDOV ‡([SORUHUHODWLRQVKLSEHWZHHQVRQJDQGPDOHILWQHVVE\DQDO\]LQJQHVWVXFFHVVDQG WHUULWRU\TXDOLW\ )XWXUH:RUN $QDO\VHV ‡1DUDQJR'/5RGHZDOG$'8UEDQDVVRFLDWHGGULYHUVRIVRQJYDULDWLRQDORQJDUXUDO±XUEDQ JUDGLHQW %HKDYLRUDO(FRORJ\ GRLGRLEHKHFRDUY ‡6DOHVV5$FRXVWLFVWUXFWXUHVLQJLQJEHKDYLRUDQGYRFDOSHUIRUPDQFHWUDGHRIIVLQWKH1RUWKHUQFDUGLQDO &DUGLQDOLVFDUGLQDOLV )$80DVWHUV7KHVLV3URSRVDO ‡6SLHU67UDIILFQRLVHDQGVH[XDOVHOHFWLRQVWXGLHVRIDQWKURSRJHQLFLPSDFWRQELUGVRQJVDQG XQGHUJUDGXDWHVWXGHQWUHDVRQLQJRIHYROXWLRQDU\PHFKDQLVPV81/0DVWHUV7KHVLV 5HIHUHQFHV )LJXUH6SHFWURJUDPRIVRQJVXQJE\PDOH1<<5&)6UXUDOFRPSRVHGRI WZRWULOOV7ULOO$DQG7ULOO% 7LPHVHFRQGV )UHTXHQF\N+]



PAGE 1

y = 0.4198x + 18.511 R² = 0.0335 R = 0.1830 0 5 10 15 20 25 30 35 0 1 2 3 4 5 6 7 Wet Weight of Above Hinge Region (g) Ratio of Y/X 0 0.5 1 1.5 2 2.5 Small Medium Large Hinge Length (cm) Leaf Size 0 0.5 1 1.5 2 2.5 3 3.5 Small Medium Large Lenght of Below Hinge Region (cm) Leaf Size 0 5 10 15 20 25 Small Medium Large Y/X Leaf Size 0 0.1 0.2 0.3 0.4 0.5 0.6 D1 D2 D3 D4 D5 D6 Diameter (cm) Petiole Position Small Medium Large Results 1 2 3 4 5 6 A Morphometrics Study of Prostrate Leaves of Wintergreen Fern Dryopteris intermedia Leaf Length Ranges (cm) Small 16.8 40.6 Medium 41.5 59.5 Large 60.1 78.3 Wintergreen fern Dryopteris intermedia (Intermediate Wood Fern) is a common understory herbaceous fern that inhabits much of the Eastern United States and Canada (Nielsen 2017). In early winter leaves become prostrate, forming a hinge in the proximal region of the petiole that allows them to lay flat. In the temperate regions of upstate New York, being wintergreen has several advantages. Wintergreen ferns have a prolonged photosynthetic period, while also retaining valuable nutrients, allowing them to thrive in nutrient poor environments ( Aerts 1995). Many studies have been published on the effects of prostration on wintergreen ferns, showing that without, processes such as photosynthesis and respiration would not be as efficient during the winter months (Forget et al. 2018). Prostration gives these leaves an advantage, allowing them to reach conditions optimal for photosynthesis and facilitating respiration in the airspace below (when under snow cover) (Forget et al. 2018). Although this species of fern is well known, there has not been previous studies regarding the morphometrics of the hinge formation on the petiole. Introduction Prostrated leaves from six different clones of similar size located on Blue Trail at Rice Creek Field Station were collected in early winter. From each clone, three leaves in each size group were collected (small , medium , and large sized leaves; see table below ). The weight of different regions of the fresh leaves were measured within two hours of collection. These measurements included, the length and fresh leaf weight of the region of the petiole above the hinge, the length and fresh weight of the region of the petiole below the hinge, the entire length of the leaf, length of the hinge region and the diameter of six points along the entire petiole of the leaf. These measurements are illustrated in Figure 1. Material and Methods Figure 1: This figure represents the several points of measurement on the leaf. Figure 2: Average hinge length for the small , medium , and large sized leaves. Figure 3: Average length of the below hinge region for small , medium , and large sized leaves Figure 4: Average ratio of the above hinge region over the below hinge region for small , medium , and large sized leaves. Figure 5: Diameter measurements at six points along the petiole, for small , medium, and large sized leaves. Figure 6: Linear regression of fresh weight of the above hinge zone versus the relative position of the hinge (Y/X). For figures: Triple asterisk indicates significance level of P < .001. Double asterisks indicates significance level of p < .005. Single asterisks indicates significance level of P < .05. Results: Figures 1 6 We hypothesize that weight may contribute to where the hinge forms. What is the average hinge length for each of the three sizes? What is the average distance of the hinge from the bottom of the petiole for all three size groups? Is there a significant difference between the position of the hinge for all three size groups? What is the diameter at different points on the petiole, and do the diameters suggest stronger mechanical support in some regions of the leaf? Dose the position of the hinge correlate to the weight of the above hinge region? Objectives Previous research indicated that the length of the zone where the hinge forms is 1 2 cm for Polystichum acrostichoides ( Nooden and Wagner, 1997). Our research indicates the hinge zone is much more variable: 1 6 cm. Weight might be a potential cause for the relative location of the hinge, but more experimental studies are needed to conclude that weight plays a role in the relative location of the hinge. In conclusion, weight might be the deciding factor of the relative location of the hinge zone on the petiole, and this location might be the balance point between the weight of the leaf and the gradually increasing level of mechanical support towards the base of the petiole. Discussion * *** *** ** *** *** *** *** *** *** *** *** Robert Salerno and Jinyan Guo Department of Biological Sciences



PAGE 1

Regulation of Ras Associated Protein 1 By Kinase Responsive to Stress B in Dictyostelium discoideum Tiffany Flores, Dr. Yulia Artemenko Department of Biological Sciences, SUNY Oswego, Oswego, NY This work was supported by NSF RUI grant no. 1817378 (to Y.A.). Acknowledgements CLONING : ! PCR was successful for Rap1 and digest of both the insert and vector were successful. Gel extraction also proved to work for Rap1, and two different ligations were used to do transformation. ! Colonies from transformation were screened and gave the expected pattern in the diagnostic digest. ! Positive clones were confirmed by sequencing. WESTERN BLOT: ! Immunoblotting showed that the antibody against mCherry can be used to detect RFP tagged Rap1. ! Rap1 (G12V) in KrsB null cell lysates and wild type cell lysates was detected on immunoblots, but did not show the electrophoretic shift under the current conditions. Future Directions ! Analyze electrophoretic mobility of RFP tagged Rap1 or Rap1 G12V in KrsB null and wild type cells following stimulation of cells with a chemoattractant. ! Analyze Rap1 Ð RFP localization in KrsB null and wild type cells by confocal microscopy. Dictyostelium discoideum is a social amoeba that is commonly used as a model organism for studying chemotaxis, which is a directed migration along a chemical gradient, due to its similarities to human neutrophils and metastatic cancer cells. There are multiple pathways involved in regulating migration. In particular, kinase responsive to stress B (KrsB), a homolog of mammalian tumor suppressor MST1/2 and Drosophila Hippo, is a negative regulator of cell adhesion and migration in D. discoideum. However, little is known about the molecular mechanism of KrsB action. Another regulator of adhesion is small GTPase Ras associated protein 1 (Rap1), which acts by affecting talin and myosin II. In mammalian cells Rap1 can be phosphorylated, which leads to its inhibition. We hypothesized that KrsB might negatively regulate Rap1 by phosphorylation, thereby disrupting the activation of Rap1 on the membrane. To determine if KrsB phosphorylates Rap1 we will perform immunoblotting for Rap1 in cells with or without KrsB and look for a shift in the electrophoretic mobility as an indicator of phosphorylation. In this study, we were able to detect RFP tagged constitutively active Rap1 G12V on an immunoblot using an antibody against mCherry. We will now continue to conduct immunoblotting to detect mobility shifts of phosphorylated Rap1. To be able to track Rap1 localization, we successfully generated an RFP Rap1 expression construct. We will examine RFP Rap1 localization in cells with or without KrsB. Abstract ! Dictyostelium discoideum is a social amoeba that has been used widely as a model organism. ! The life stages of Dictyostelium discoideum make it uniquely suited to study migration and chemotaxis Ð processes that contribute to human diseases such as cancer. 1 ( https://en.wikipedia.org/wiki/Dictyostelium_discoideum ) KrsB (Kinase Responsive to Stress B) ! KrsB is a homolog of tumor suppressors MST1/2 in mammalian cells and Drosophila Hippo. 1 ! KrsB plays a role in chemotaxis by being a negative regulator of cell adhesion and migration. 1 ! Cells lacking KrsB have increased contact with the surface and are more difficult to detach. 1 Rap1 (Ras Associated Protein 1) ! Rap1 is a small GTPase that is known to regulate adhesion in mammalian cells and in Dictyostelium discoideum. 2,4 ! In mammalian cells, Rap1 can be regulated by phosphorylation by cAMP dependent protein kinase A (PKA), and this phosphorylation negatively regulates Rap1. 2 ! Previous studies showed that Rap1 can regulate cell adhesion without KrsB, although KrsB might modulate Rap1 function. 4 HYPOTHESIS ! KrsB negatively regulates Rap1 by phosphorylation, which disrupts activation of Rap1 on the membrane. Introduction Approach To look at the regulation of Rap1 by KrsB we plan to: 1) Look at the localization of RFP tagged Rap1 in the presence or absence of KrsB ! This requires cloning of Rap1 into an expression plasmid with an RFP gene. 2) Look at the phosphorylation of Rap1 in the presence or absence of KrsB ! To do this we need to examine if RFP tagged Rap1 has an electrophoretic mobility shift indicative of phosphorylation in cells with KrsB compared to without. ! To perform immunoblotting for RFP tagged Rap1, first we need to test whether the mCherry antibody is able to recognize the RFP tag. Methodology Discussion References 1. Artemenko, Y., Batsios, P., Borleis, J., Gagnon, Z., Lee, J., Rohlfs, M., Sanseau, D., Willard, S., Schleiher, M., & Devreotes, P. (2012). Tumor suppressor Hippo/MST1 kinase mediates chemotaxis by regulating spreading and adhesion. PNAS , 13632 13637. doi: 10.1073/pnas.1211304109. 2. Takahashi M, Dillon TJ, Liu C, Kariya Y, Wang Z, Stork PJ. Protein kinase A dependent phosphorylation of Rap1 regulates its membrane localization and cell migration. J Biol Chem . 2013;288(39):27712 27723. doi:10.1074/jbc.M113.466904 3. DictyBase. Dictyostelium discoideum : Model System in Motion. http://dictybase.org/tutorial/ (accessed August 17, 2020) 4. Niu , G. (2020). Genetic Interaction between Adhesion Regulators Rap1 and Kinase Responsive to Stress B in Dictyostelium discoideum. Honors Thesis, SUNY Oswego. Cloning of RFP Rap1 Figure 1. Diagnostic digest of plasmid with the Rap1 gene. Diagnostic digest was conducted for plasmid DNA isolated from bacterial colonies after transformation. The plasmid was digested with EcoRI and HindIII . The wells on the top panel showed the expected band sizes at around 4838 bp, 2411 bp, and 837 bp. Figure 2. Detection of RFP tagged constitutively active Rap1 (G12V) with an antibody against mCherry . Wild type cells expressing RFP Rap1 G12V were lysed, proteins were separated by SDS PAGE and transferred to a PVDF membrane. Membrane was immunoblotted with a primary antibody against mCherry , followed by a secondary antibody conjugated with horseradish peroxidase. Signal was detected by chemiluminescence. Signal was detected at the expected size (between 55 to 40 kDa ) !" Results Results Figure 3. Detection of RFP tagged constitutively active Rap1 (G12V) in KrsB null and wild type cells with an antibody against mCherry. Wild type and KrsB null cells expressing RFP Rap1 G12V were lysed, proteins were separated by SDS PAGE and transferred to a PVDF membrane. Membrane was immunoblotted with a primary antibody against mCherry, followed by an anti rabbit secondary antibody. Signal was detected by chemiluminescence.