The ENCODE program is the post-genomics program initiated by National Human Genome Research Institute to annotate the human genome. We use genome tiling microarrays to determine origins of replication, replication profile, replication factor binding sites and epigenetics of the human genome. Obviously the next step will be to see how these are changed in cancers. (http://genome.bioch.virginia.edu/encode)
that stems from an imbalance of replication factors
that stems from abnormalities of DNA replication or from external agents such as chemotherapeutic agents or radiation. The response pathways often use tumor suppressor proteins like p53, Fanconi Anemia complex or BRCA1. A related area of research focuses on two putative AAA+ ATPases (Rvb1 and 2) that are part of chromatin-remodeling complexes that respond to DNA damage.
(widely used clinically) and their utility in suppressing the cancer cell-cycle.
We are examining how the new "dark matter" of the genome, microRNAs, regulate cell proliferation and differentiation in muscle differentiation and in prostate cancer cells.
Principal Investigator
Research Assistant Professor
Yuichi J. Machida (Ph.D. Nagoya University, Nagoya)
Postdocs
Tarek Abbas (Ph.D. CUNY, New York)
Sudhakar Jha (Ph.D. JNU, New Delhi)
Neerja Karnani (Ph.D. JNU, New Delhi)
Yong Sun Lee (Ph.D. Seoul National University, Seoul)
Sara Hook (Ph.D. Duke University, Durham)
Yoshiyuki Shibata (Ph.D. Kyushu University, Fukuoka)
Uma Sivaprasad (Ph.D. Ohio State University, Columbus)
Kenta Terai (Ph.D. Osaka University, Osaka).
Sukumar Sarkar (Ph.D. Kobe University, Kobe)
Research Assistant
Yuka Machida
Graduate Students
Ankit Malhotra (Ph.D. student, Biochemistry, M.C.S., Computer
Science)
Christopher Taylor (Ph.D. student, M.S. Computer Science)
Jamie Teer (Ph.D. student, BBS, Harvard)
Jie Lin (Ph.D. student, Biochemistry)
Adam Mueller (MD, Ph.D. student, Biochemistry)
Undergraduate Students
Somsakul Wongpalee (UVa)
Charles Dyer (UVa)
Staff
Nancy Rush
Alumni
Junjie Chen, Professor, Yale Medical School
Partha Saha, Sr. Lecturer, Saha Institute, Kolkata
Suman K. Dhar, Assoc. Professor, JNU, New Delhi
Zophonias Jonsson, Sr. Lecturer, University of Iceland
David Garcia Quintana, Asst. Professor, Autonomous University, Barcelona.
Chinweike Ukomadu, Asst. Professor, Harvard Medical School
Yi Ling Lin, Asst. Professor, U. of Kentucky
James Wohlschlegel, Asst. Professor, UCLA
Sandeep Saxena, Asst. Professor, National Institute of Immunology, New Delhi
Takeshi Senga, Assoc. Professor, Nagoya University
Kenichi Yoshida, Assoc. Professor, Meiji University
Yuefeng Chen, Research Assistant, Eli Lilly, Indianapolis
A. Original Articles in Refereed Journals
- Dutta A, Wang L-H, Hanafusa T, Hanafusa H. Partial nucleotide sequence of Rous sarcoma virus-29 provides evidence that the original Rous sarcoma virus was replication defective. J Virol. 1985; 55, 728-735.
- Dutta A, Majumder H K. Effects of monosaccharides on uptake of Leishmania donovani promastigotes by murine macrophages. Indian Jl Biochem Biophys. 1986; 23, 1-4.
- Dutta A, Dorai T, Hanafusa H. The putative trans-activator in the MA gag region of Rous sarcoma virus is not required for cell transformation. J Virol. 1988; 62, 4767-4769.
- Prywes R, Dutta A, Cromlish J A, Roeder R G. Phosphorylation of serum response factor, a factor that binds to the serum response element upstream of the c-fos promoter. Proc Natl Acad Sci USA. 1988; 85, 7206-7210.
- Dutta A, Stoeckle M Y, Hanafusa H. Serum and v-src increase the level of CCAAT binding factor required for transcription from a retroviral LTR. Genes & Dev. 1990; 4, 243-254.
- Dutta A, Hamaguchi M, Hanafusa H. Serum independence of transcription from the promoter of an avian retrovirus in v-src transformed cells is a primary, intracellular effect of increased tyrosine phosphorylation. Proc Natl Acad Sci U S A. 1990; 87, 608-612.
- Dutta A, Stillman B. cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication. EMBO J. 1992; 11, 2189-2199.
- Dutta A, Ruppert J M, Aster J C, Winchester E. Inhibition of DNA replication factor RPA by p53. Nature. 1993; 365, 79-82. (Accompanied by News and Views piece in Nature)
- Chen J, Jackson P K, Kirschner M W, and Dutta A. Separate domains of p21 involved in the inhibition of cdk kinase and PCNA. Nature. 1995; 374, 386-388
- Dutta A, Chandra R, Leiter L M, Lester S. Cyclins as markers of tumor proliferation and angiogenesis: immunocytochemical studies in breast cancer. Proc Natl Acad Sci USA. 1995; 92, 5386-5390.
- Keshav K F, Chen C, Dutta A. Rpa4, a homolog of the 34 kDa subunit of the replication protein A complex. Mol. Cell. Biol. 1995; 15, 3119-3128.
- Chen J, Peters R, Saha P, Lee P, Theodoras A, Pagano M, Wagner G, Dutta A. A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo. Nucleic Acids Research. 1996; 24, 1727-1733.
- Lin Y-L, Chen C, Keshav K F, Winchester E, Dutta A. Dissection of functional domains of the human DNA replication protein complex Replication Protein A. J. Biol. Chemistry. 1996; 271, 17190-17198.
- Leiter L M, Chen J, Marathe T, Tanaka M, Dutta A. Loss of transactivation and transrepression function, and not RPA binding, alters growth suppression by p53. Oncogene. 1996; 12, 2661-2668.
- Henricksen L A, Carter T, Dutta A, Wold M S. Phosphorylation of human replication protein A by the DNA-dependent protein kinase is involved in the modulation of DNA replication. Nucleic Acids Research. 1996; 24, 3107-3112.
- Chen J, Saha P, Kornbluth S, Dynlacht B, Dutta A. Cyclin binding motifs are essential for the function of p21/CIP1. Mol. Cell. Biol. 1996; 16, 4673-4682.
- Mashal R D, Lester S, Corless C, Richie J P, Chandra R, Propert K J, Dutta A. Expression of cell-cycle regulated proteins in prostate cancer. Cancer Research. 1996; 56, 4159-4163.
- Chen J, Chen S, Saha P and Dutta A. p21 disrupts the recruitment of human Fen1 by proliferating cell nuclear antigen into the DNA replication apparatus. Proc. Natl. Acad. Sci. USA. 1996; 93, 11597-11602.
- Saha P, Eichbaum Q, Silberman E D, Mayer B J, Dutta A. p21/CIP1 and Cdc25A, competition between an inhibitor and an activator of cyclin dependent kinases. Mol. Cell. Biol. 1997; 17, 4338-4345.
- Quintana DG, Hou ZH, Thome KC, Hendricks M, Saha P, Dutta A. Identification of a novel subunit of the human origin recognition complex with homology to yeast Orc4. J. Biol. Chemistry 1997; 272, 28247-28251.
- Lin Y L, Shivji M K K, Chen C, Kolodner R, Wood R D, Dutta A. The evolutionarily conserved zinc finger motif in the largest sub-unit of human RPA is required for DNA replication and mismatch repair but not for nucleotide excision repair. J. Biol. Chemistry 1998; 273, 1453-1461.
- Saha P, Chen J, Thome K C, Lawlis S J, Hou Z H, Hendricks M, Parvin J D, Dutta A. The human CDC6/Cdc18 associates with Orc1 and cyclin-cdk and is selectively eliminated from the nucleus at the onset of S phase. Mol. Cell. Biol. 1998; 18, 2758-2767.
- Saha P, Thome K C, Yamaguchi R, Hou Z H., Weremowicz S and Dutta A. The human homolog of Saccharomyces cerevisiae CDC45. J. Biol. Chemistry 1998; 273, 18205-18209.
- Renshaw A A, Loughlin K R, Dutta A. Cyclin A and Mib1 (Ki67) as markers of proliferation in primary renal neoplasms. Modern Pathology. 1998; 10, 963-966.
- Quintana DG, Thome KC, Hou ZH, Ligon AH, Morton C C and Dutta A. ORC5L, a new member of the human Origin Recognition Complex, is deleted in uterine leiomyomas and malignant myeloid diseases. J. Biol. Chemistry 1998; 273, 27137-27145.
- Qiu X B, Lin Y L, Thome K C, Pian P, Schlegel B P, Weremowicz S, Parvin J D, Dutta A. An Eukaryotic RuvB-like Protein (RUVBL1) Essential for Growth. J. Biol. Chemistry 1998; 273, 27786-27793.
- Quade B J, Park J J, Crum C P, Sun D, Dutta A. In vivo cyclin E expression as a marker for early cervical neoplasia. Modern Pathology. 1998; 11, 1238-1246.
- Pinto S, Quintana D G, Smith P, Mihalik R M, Hou Z -H, Boynton S, Jones C J, Hendricks M, Velinzon K, Wohlschlegel J A, Austin R J, Lane W S, Tully T* and Dutta A*. latheo encodes a subunit of the Origin Recognition Complex and disrupts neuronal proliferation and adult olfactory memory when mutant. Neuron. 1999; 23, 45-54. * Co-corresponding authors.
- Datta M W, Renshaw A A, Dutta A, Hoffman M A and Loughlin KR. Evaluation of cyclin expression in testicular germ cell tumors: cyclin E correlates with tumor type, advanced clinical stage, and pulmonary metastasis. Modern Pathology. 2000; 13, 667-672
- Dhar S K and Dutta A. Identification of the human ORC6 homolog. J. Biol. Chemistry. 2000; 275, 34983-34988..
- Thome K C, Dhar S K, Quintana D G, Delmolino L, Shahsafaei A and Dutta A. Subsets of human ORC subunits are expressed in non-proliferating cells and associate with non-ORC proteins. J. Biol. Chemistry. 2000; 275, 35233-35241.
- Yamaguchi R and Dutta A. Proteasome inhibitors alter the orderly progression of DNA synthesis in S phase of HeLa cells and lead to re-replication of DNA. Exptl. Cell Research. 2000; 261, 271-283.
- Wohlschlegel J A, Dwyer B, Dhar S K, Cvetic C, Walter J and Dutta A. Inhibition of eukaryotic DNA replication by geminin binding to Cdt1. Science. 2000; 290, 2309-2312. (Accompanied by Perspectives essay in Science)
- Takeda D, Wohlschlegel J A, and Dutta A. A bipartite substrate recognition motif for cyclin-dependent kinases. J. Biol. Chemistry. 2001; 276, 1993-1997.
- Jonsson Z O, Dhar S K, Narlikar G, Auty R, Wagle N, Pellman D, Pratt R E, Kingston R and Dutta A. Rvb1p and Rvb2p are essential components of a chromatin remodeling complex that regulates transcription of over 5% of yeast genes. J. Biol. Chemistry. 2001; 276,16279-16288.
- Yoshida K, Kuo F, George E L, Sharpe A H and Dutta A. Requirement of CDC45 for postimplantation mouse development. Mol. Cell. Biol. 2001; 21, 4598-4603.
- Wohlschlegel J A, Dwyer B, Takeda D and Dutta A. A mutational analysis of the Cy motif from p21 reveals sequence degeneracy and specificity for different cyclin-dependent kinases. Mol. Cell. Biol. 2001; 21, 4868-4874.
- Delmolino L M, Saha P and Dutta A. Multiple mechanisms regulate subcellular localization of human CDC6: NLS, NES and phosphorylation. J.Biol. Chemistry. 2001; 276, 26947-26954.
- Dhar S K, Delmolino L M and Dutta A. Architecture of the human Origin Recognition Complex. J Biol. Chemistry, 2001; 276, 29067-29071.
- Dhar S K, Yoshida K, Machida Y, Khaira P, Chaudhuri B, Wohlschlegel J A, Leffak M, Yates J and Dutta A. Replication from oriP of Epstein-Barr Virus requires human ORC and is inhibited by geminin. Cell, 2001; 106, 287-296.
- Chaudhuri B, Xu H, Todorov I, Dutta A and Yates J L. Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus. . Proc Natl Acad Sci U S A. 2001; 98, 10085-10089.
- Wohlschlegel J A, Dhar SK, Prokhorova TA, Dutta A* and Walter J*. Xenopus Mcm10 binds to origins of DNA replication after Mcm2-7 and stimulates binding of Cdc45. Molecular Cell. 2002; 9, 233-240. * Co-corresponding authors. (F1000)
- Wohlschlegel J A, Kutok J L , Weng A P and Dutta A. Expression of geminin as a marker of cell proliferation in normal tissues and malignancies. Am. J. Pathology 2002; 161, 267-273.
- Ukomadu C and Dutta A. Inhibition of cdk2 activating phosphorylation by mevastatin. J. Biol. Chemistry 2003; 278, 4840-4846.
- Vaziri C, Saxena S, Jeon Y, Lee C, Murata K, Machida Y, Wagle N, Hwang D S and Dutta A. A p53 dependent checkpoint pathway prevents re-replication. Molecular Cell 2003; 11, 997-1008. (Cover Image)
- Ukomadu C and Dutta A. p21 dependent inhibition of colon cancer cell growth by mevastatin is independent of inhibition of G1 cyclin-dependent kinases. J. Biol. Chem. 2003; 278, 43586
- Saxena S, Jonsson Z O and Dutta A. Small RNAs with imperfect match to endogenous mRNA repress translation: implications for off-target activity of siRNA in mammalian cells. J. Biol. Chem. 2003; 278, 44312-44319.
- Yoshida K, Oyaizu N, Dutta A and Inoue I. The destruction box of human Geminin is critical for proliferation and tumor growth in human colon cancer cells. Oncogene. 2004; 23, 58-70.
- Zhu W, Chen Y and Dutta A. Re-replication by depletion of geminin is seen regardless of p53 status and activates a G2/M checkpoint. Mol. Cell. Biol. 2004; 24, 7140-7150. (Cover Image)
- Saxena S, Yuan P, Dhar SK, Senga T, Takeda D, Robinson H, Kornbluth SA, Swaminathan K and Dutta A. A dimerized coiled-coil domain and an adjoining part of geminin interact with two sites on Cdt1 for replication inhibition. Molecular Cell, 2004; 15, 245-258.
- Jónsson Z O, Jha S, Wohlschlegel J A and Dutta A. Rvb1p/Rvb2p recruit Arp5p and assemble a functional Ino80 chromatin remodeling complex. Molecular Cell, 2004; 16, 465-477.
- ENCODE Project Consortium. The ENCODE (ENCyclopedia Of DNA Elements) Project. Science, 2004;306, 636-640.
- Lee YS, Kim HK, Chung S, Kim KS, and Dutta A. Depletion of human microRNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation. J Biol Chem. 2005; 280, 16635-16641.
- Jeon Y, Bekiranov S, Karnani N, Kapranov P, Ghosh S, MacAlpine D, Lee C, Hwang DS, Gingeras T and Dutta A. Temporal profile of replication of human chromosomes. Proc. Natl. Acad. Sci. USA. 2005; 102, 6419-6424.
- Takeda D Y, Parvin J D and Dutta A. Degradation of Cdt1 during S phase is SKP2 independent and is required for efficient progression of mammalian cells through S phase. J. Biol. Chem. 2005; 280, 23416-23423.
- Machida Y J, Teer J K and Dutta A. Acute reduction of an ORC subunit in human cells reveals a requirement of ORC for CDK2 activation. J. Biol. Chem. 2005; 280, 27624-27630.
- Takeda D Y, Shibata Y, Parvin J D and Dutta A. Recruitment of ORC or CDC6 is sufficient to create an artificial origin of replication in mammalian cells. Genes & Development. 2005; 19, 2827-36. (Research Highlight in Nature Reviews Genetics and Nature Reviews MCB; F1000)
- Senga T, Sivaprasad U, Zhu W, Park J H, Arias E E, Walter J C and Dutta A. PCNA is a co-factor for Cdt1 degradation by CUL4/DDB1 mediated N-terminal ubiquitination. J. Biol. Chem. 2006; 281, 6246-52. (F1000)
- Teer J K, Machida Y J, Labit H, Novac O, Hyrien O, Marheineke K, Zannis-Hadjopoulos M and Dutta A. Proliferating human cells hypomorphic for Orc2 and pre-RC formation have a defect in p53 activation and CDK2 kinase activation. J. Biol. Chem. 2006; 281, 6253-60.
- Ernkvist M, Aase K, Ukomadu C, Wohlschlegel J, Blackman R, Veitonmaki N, Bratt A, Dutta A and Holmgren L p130-Angiomotin associates with actin and controls endothelial cell shape. FEBS Journal, 2006; 273, 2000-11
- Zhu W and Dutta A. An ATR, BRCA1 mediated Fanconi Anemia pathway is required for activating the G2/M checkpoint and DNA damage repair upon re-replication. Mol. Cell. Biol. 2006; 26, 4601-11.
- Kim HK, Lee YS, Sivaprasad U, Malhotra A and Dutta A. Muscle-specific microRNA miR-206 promotes muscle differentiation. J. Cell Biol. 2006; 174, 677-687. (Cover Image and News Highlight in JCB; Research Highlight in Nature, 2006: 443, 4-5; F1000)
- Machida Y J, Machida Y, Chen Y, Gurtan A, Kupfer G M, D’Andrea A D and Dutta A. UBE2T is the E2 in the Fanconi Anemia Pathway and Undergoes Negative Autoregulation. Molecular Cell, 2006; 23, 589-96.
- Sivaprasad U, Abbas T and Dutta A. Differential efficacy of HMG CoA reductase inhibitors on the cell cycle of prostate cancer cells. Molecular Cancer Therapeutics, 2006; 5: 2310-6.
- Machida Y J, Chen Y, Machida Y, Malhotra A, Sarkar S and Dutta A. Targeted Comparative RNAi (TARCOR) Analysis Reveals Differential Requirement of Genes Essential for Cell Proliferation. Mol. Biol. Cell. 2006; 17: 4837–4845.
- Karnani N, Taylor C M, Malhotra A and Dutta A. Pan-S replication patterns and chromosomal domains defined by genome tiling arrays of ENCODE genomic areas. Genome Research, 2007; 17 : 865-76.
- Machida YJ and Dutta A. The APC/C Inhibitor, Emi1, Is Essential for Prevention of Rereplication. Genes & Development. 2007; 21:184-194.(Cited by F1000)
- Abbas T, Jha S, Sherman NE and Dutta A. Autocatalytic Phosphorylation of CDK2 at the Activating Thr160. Cell Cycle, 2007; 6:843-52. (F1000 citation)
- Lee YS and Dutta A. The tumor suppressor microRNA let-7 represses the HMGA2 oncogene. Genes & Development, 2007; 21:1025-30. (Accompanied by a Perspectives essay).
- ENCODE project consortium. Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project consortium. Nature, 2007: 799-816. (Our group provided and analyzed the DNA replication data for the consortium.)
- Zhu W, Ukomadu C, Jha S, Senga T, Dhar SK, Wohlschlegel JA, Nutt LK, Kornbluth S and Dutta A. Mcm10 and And-1/CTF4 recruit DNA polymerase to chromatin for initiation of DNA replication. . Genes & Development. 2007; in press.
B. Reviews, Chapters and editorials
- Dutta A, Din S U , Brill S J, Stillman B. Phosphorylation of Replication Protein A: a role for cdc2 kinase in G1-S regulation. in The Cell Cycle, Cold Spring Harbor Symposium of Quantitative Biology. 1991; 56, 315-324.
- Stillman B, Bell S P, Dutta A, and Marahrens Y. Studies on DNA replication and the cell cycle. in Regulation of the eukaryotic cell cycle, CIBA Foundation, Symposium 170, Wiley Publishers, Chichester. 1992; 147-160.
- Dutta A. Trans-plication factors. Current Biology. 1993; 3, 709-712.
- Dutta A and Winchester E. SV40 based in vitro DNA replication assay. in "Cell Cycle: Materials and Methods", ed. Michelle Pagano. Springer-Verlag. Heidelberg. 1995; 175-185.
- Dutta A and Bell S P. Initiation of DNA replication in eukaryotic cells. Annual Review of Cell and Developmental Biology. 1997; 13, 293-332.
- Dutta A. Regulation of S phase. in "Cell Cycle Control", ed. Michelle Pagano. Springer-Verlag. Heidelberg. 1998; 35-56.
- Lin, Y-L and Dutta A. In vitro DNA replication. in “Current Protocols in Cell Biology”, ed. Mary Dasso. John Wiley and Sons. New York. 1998; Chapter 11, Unit 11.5.
- Dutta A. Origin Recognition Complex and other eukaryotic initiator proteins. in “Experimental Medicine” special issue on DNA replication, ed. Takuya Ohashi and Akio Matsukage. Yodosha Co. Ltd. Tokyo. 1998; 16, 53-60.
- Quintana D.G. and Dutta A. The metazoan origin recognition complex. Frontiers in Biosciences. 1999; 4, 805-815.
- Bell S P. and Dutta A. Initiation of DNA replication in eukaryotic cells. Annual Review of Biochemistry. 2002; 71, 333-374.
- Wohlschlegel J A, Dutta A and Dhar S K. ORC and the initiation of DNA replication. In Eukaryotic DNA Replication for ChemTracts - Biochemistry & Molecular Biology; ed M. L. Depamphilis. 2002; 15, 533-543.
- Saxena S. and Dutta A. Geminin-Cdt1 balance is critical for genetic stability. In Mutation Research: Fundamental and Molecular Mechanisms of Mutagenesis- Stress Response; ed Nic Denko and Al Fornace, 2005; 569, 111-121.
- Machida Y J and Dutta A. Cellular checkpoint mechanisms monitoring proper initiation of DNA replication. J. Biological Chemistry, 2005; 280, 6253-6256.
- Blow J A and Dutta A. Preventing re-replication of chromosomal DNA. Nature Reviews Molecular Cell Biology, 2005; 6, 476-486.
- Takeda D and Dutta A. DNA replication and progression through S phase. Oncogene, 2005; 24, 2827-2843.
- Zhu W, Abbas T and Dutta A. DNA replication and genomic instability. "Genome instability and cancer development", ed. Erich Nigg, Series: Advances in Experimental Medicine and Biology, 2005; 570, 249-279 .
- Karnani N, Dutta A. Nuclear localization of RFC40 by RIalpha: a link between cellular signaling and proliferation. Cancer Biol Ther. 2005; 4, 438-9.
- Teer, J.K. and Dutta, A. (2006) Regulation of S Phase. In: Kaldis, P. (ed.) Cell Cycle Regulation. Results and Problems in Cell Differentiation, vol. 42. Springer Berlin, Heidelberg, New York, pp 31-64.
- Machida YJ, Hamlin JL and Dutta A. Right place, right time, and only once: replication initiation in metazoans. Cell. 2005; 123, 13-24.
- Sivaprasad U, Dutta A and Bell SP. Assembly of Pre-replication Complexes. “In DNA replication and human disease (ed. M.L. DePamphilis), pp. 141-154. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. 2006; in press.
- Abbas T and Dutta A. CDK2-Activating Kinase (CAK): More Questions than Answers. Cell Cycle. 2006; 5, 1123-1124.
- Lee YS and Dutta A. MicroRNAs: small but potent oncogenes or tumor suppressors. Curr. Opinion in Investigational Drugs 2006; 7, 560-564.
- Zhu W and Dutta A. Activation of fanconi anemia pathway in cells with re-replicated DNA. Cell Cycle. 2006; 5, 2306-2309
- Dutta A. Chaotic license for genetic instability and cancer. Nature Genetics, 2007; 39, 10-11.
- Sivaprasad U, Machida YJ and Dutta A. APC/C - the master controller of origin licensing? Cell Division, 2007; 2, 8.
The slides for class "MICR811" can be found here
Some photos from "Lab Barbecue 2007" can be found here
Department of Biochemistry and Molecular Genetics,
Room no. 1232, Jordan Hall,
University of Virginia,
Charlottesville, VA 22908
Ph: 434 - 924 - 2466