John 'Jay' Schneekloth Jr., Ph.D.
John 'Jay' Schneekloth Jr., Ph.D.
Investigator
Head, Chemical Genetics Section

My laboratory uses chemical biology approaches to identify and study small molecule probes of gene expression. We are interested in two areas: protein sumoylation (a ubiquitin-like posttranslational modification that regulates transcription factor activity) and RNA-binding small molecules. We use synthetic chemistry and high throughput screening techniques to identify natural products or synthetic compounds that inhibit targets of interest.

Link to additional information about Dr. Schneekloth’s research.

Areas of Expertise
1) organic chemistry, 2) natural products, 3) chemical biology, 4) ubiquitin-like signaling, 5) RNA-binding small molecules

Contact Info

John 'Jay' Schneekloth Jr., Ph.D.
Center for Cancer Research
National Cancer Institute
Building 376, Room 225C
Frederick, MD 21702-1201
301-228-4620
schneeklothjs@mail.nih.gov

Small Molecule Probes of Protein Sumoylation

The Small Ubiquitin-like Modifier (SUMO) is a ubiquitin-like posttranslational modification. Protein sumoylation is tightly linked with gene expression, as many of the known substrates for sumoylation are transcription factors. Furthermore, the aberrant regulation of sumoylation is linked to a number of cancers. Our laboratory has developed a novel electrophoretic mobility shift assay we are using in a high throughput screen of natural product extracts to identify naturally occurring small molecule sumoylation inhibitors. As a complementary approach, we are using fragment-based inhibitor design approaches to develop synthetic inhibitors of sumoylation. In addition, we have developed several synthetic substrates of sumoylation with the aim of gaining insight into substrate recognition and to identify active site-directed inhibitors. A major goal of this project is to establish a structural basis for chemical inhibition of sumoylation, and to gain insight into the structure and function of sumoylation enzymes and their role in cancer biology.

Identification of RNA- and DNA-binding Small Molecules Using Small Molecule Microarrays

The identification of selective RNA- and DNA-binding small molecules has been a longstanding challenge for chemical biology. Our approach to this problem is to use small molecule microarrays as a screening technology. We have assembled a library of 20,000 compounds that are used to screen diverse nucleic acid targets such as RNA hairpins and DNA G-quadruplexes. Most recently, we reported the identification of a druglike compound that binds to the HIV transactivation response (TAR) hairpin. This compound is not cationic, selectively binds to TAR in the context of the entire HIV 5'UTR, and rescues lymphoblastic cells from HIV-mediated cytopathicity without any observable toxicity. Future goals for this project involve the identification of small molecules that selectively interact with a broad variety of therapeutically relevant RNA targets.

Scientific Focus Areas:
Cell Biology, Chemical Biology, Molecular Biology and Biochemistry
  1. Sztuba-Solinska J, Shenoy SR, Gareiss P, Krumpe LR, Le Grice SF, O'Keefe BR, Schneekloth JS.
    J. Am. Chem. Soc. 136: 8402-10, 2014. [ Journal Article ]
  2. Kim YS, Keyser SG, Schneekloth JS.
    Bioorg. Med. Chem. Lett. 24: 1094-7, 2014. [ Journal Article ]
  3. Kim YS, Nagy K, Keyser S, Schneekloth JS.
    Chem. Biol. 20: 604-13, 2013. [ Journal Article ]
  4. Kim J, Schneekloth JS, Sorensen EJ.
    Chem Sci. 3: 2849-2852, 2012. [ Journal Article ]
  5. Noblin DJ, Page CM, Tae HS, Gareiss PC, Schneekloth JS, Crews CM.
    ACS Chem. Biol. 7: 2055-63, 2012. [ Journal Article ]

Dr. Schneekloth received his undergraduate degree from Dartmouth College in 2001, where he worked with Prof. Gordon Gribble. He then moved to Yale University and obtained a Ph.D. from the chemistry department with Prof. Craig Crews in 2006. As a graduate student he studied natural product total synthesis and chemical biology relating to the ubiquitin-proteasome pathway. He then pursued an NIH postdoctoral fellowship with Prof. Erik Sorensen at Princeton University, where he worked on the development of a new multicomponent reaction and the application of this reaction to the synthesis of analgesic natural products. He returned to Yale in 2009 where he worked as a medicinal chemist at the Yale Small Molecule Discovery Center. In 2011, Dr. Schneekloth joined NCI where his research involves using synthetic chemistry and high throughput chemical biology approaches to develop chemical probes of signal transduction pathways and gene expression. Specific areas of interest include protein sumoylation (including mechanistic probes, natural products, and synthetic inhibitors) and the use of small molecule microarrays to identify RNA- and DNA-binding small molecules.

Name Position
Fardokht Abulwerdi Ph.D. Postdoctoral Fellow
Robert Boer Postdoctoral Fellow (CRTA)
David Calabrese Ph.D. Postdoctoral Fellow
Colleen Connelly Ph.D. Postdoctoral Fellow
Samuel Dahlhauser Postbaccalaureate Fellow
Will Hewitt Ph.D. Postdoctoral Fellow
Ranuka Sinniah Summer Student
Robert Wilson Summer Student
Katie Zlotkowski Ph.D. Postdoctoral Fellow

Research

Small Molecule Probes of Protein Sumoylation

The Small Ubiquitin-like Modifier (SUMO) is a ubiquitin-like posttranslational modification. Protein sumoylation is tightly linked with gene expression, as many of the known substrates for sumoylation are transcription factors. Furthermore, the aberrant regulation of sumoylation is linked to a number of cancers. Our laboratory has developed a novel electrophoretic mobility shift assay we are using in a high throughput screen of natural product extracts to identify naturally occurring small molecule sumoylation inhibitors. As a complementary approach, we are using fragment-based inhibitor design approaches to develop synthetic inhibitors of sumoylation. In addition, we have developed several synthetic substrates of sumoylation with the aim of gaining insight into substrate recognition and to identify active site-directed inhibitors. A major goal of this project is to establish a structural basis for chemical inhibition of sumoylation, and to gain insight into the structure and function of sumoylation enzymes and their role in cancer biology.

Identification of RNA- and DNA-binding Small Molecules Using Small Molecule Microarrays

The identification of selective RNA- and DNA-binding small molecules has been a longstanding challenge for chemical biology. Our approach to this problem is to use small molecule microarrays as a screening technology. We have assembled a library of 20,000 compounds that are used to screen diverse nucleic acid targets such as RNA hairpins and DNA G-quadruplexes. Most recently, we reported the identification of a druglike compound that binds to the HIV transactivation response (TAR) hairpin. This compound is not cationic, selectively binds to TAR in the context of the entire HIV 5'UTR, and rescues lymphoblastic cells from HIV-mediated cytopathicity without any observable toxicity. Future goals for this project involve the identification of small molecules that selectively interact with a broad variety of therapeutically relevant RNA targets.

Scientific Focus Areas:
Cell Biology, Chemical Biology, Molecular Biology and Biochemistry

Publications

  1. Sztuba-Solinska J, Shenoy SR, Gareiss P, Krumpe LR, Le Grice SF, O'Keefe BR, Schneekloth JS.
    J. Am. Chem. Soc. 136: 8402-10, 2014. [ Journal Article ]
  2. Kim YS, Keyser SG, Schneekloth JS.
    Bioorg. Med. Chem. Lett. 24: 1094-7, 2014. [ Journal Article ]
  3. Kim YS, Nagy K, Keyser S, Schneekloth JS.
    Chem. Biol. 20: 604-13, 2013. [ Journal Article ]
  4. Kim J, Schneekloth JS, Sorensen EJ.
    Chem Sci. 3: 2849-2852, 2012. [ Journal Article ]
  5. Noblin DJ, Page CM, Tae HS, Gareiss PC, Schneekloth JS, Crews CM.
    ACS Chem. Biol. 7: 2055-63, 2012. [ Journal Article ]

Biography

Dr. Schneekloth received his undergraduate degree from Dartmouth College in 2001, where he worked with Prof. Gordon Gribble. He then moved to Yale University and obtained a Ph.D. from the chemistry department with Prof. Craig Crews in 2006. As a graduate student he studied natural product total synthesis and chemical biology relating to the ubiquitin-proteasome pathway. He then pursued an NIH postdoctoral fellowship with Prof. Erik Sorensen at Princeton University, where he worked on the development of a new multicomponent reaction and the application of this reaction to the synthesis of analgesic natural products. He returned to Yale in 2009 where he worked as a medicinal chemist at the Yale Small Molecule Discovery Center. In 2011, Dr. Schneekloth joined NCI where his research involves using synthetic chemistry and high throughput chemical biology approaches to develop chemical probes of signal transduction pathways and gene expression. Specific areas of interest include protein sumoylation (including mechanistic probes, natural products, and synthetic inhibitors) and the use of small molecule microarrays to identify RNA- and DNA-binding small molecules.

Team

Name Position
Fardokht Abulwerdi Ph.D. Postdoctoral Fellow
Robert Boer Postdoctoral Fellow (CRTA)
David Calabrese Ph.D. Postdoctoral Fellow
Colleen Connelly Ph.D. Postdoctoral Fellow
Samuel Dahlhauser Postbaccalaureate Fellow
Will Hewitt Ph.D. Postdoctoral Fellow
Ranuka Sinniah Summer Student
Robert Wilson Summer Student
Katie Zlotkowski Ph.D. Postdoctoral Fellow