Russian Journal of Bioorganic Chemistry, Vol. 26, No. 5, 2000, р . 306

The Nuclease Activity of the Endogenous Differentiation Factor of the HL-60 Cell Line

S. M. Dranitsyna*, I. A. Kostanyan* ¹ , S. G. Andreeva*, M. V. Astapova*, I. I. Babichenko**, O. V. Baeva*, A. P. Bogachuk*, I. M. Molotkovskaya*, I. L. Rodionov***, E. V. Smirnova*, and V. M. Lipkin*

*Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences,ul. Miklukho-Maklaya 16/10, Moscow, GSP-7, 117871 Russia, ** University of Peoples ’ Friendship, ul. Miklukho-Maklaya 6, Moscow, 117198 Russia ***Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry (Pushchino Branch), Russian Academy of Sciences, Pushchino, Moscow oblast, 142292 Russia

Abstract: A structural homology between the endogenous differentiation factor of the HL-60 cell line of promyelocyte leukemia (HLDF) and several DNA/RNA-binding and DNA/RNA-hydrolyzing proteins was revealed, and expression of the hldf gene in prokaryotic systems was studied. On the basis of these experiments, the amino acid sequence of an 8-membered fragment of HLDF with potential nuclease activity was identified. The synthetic octapeptide RRWHRLKE was shown to be capable of the cleavage of RNA, linear DNA from phage , and all forms of plasmid DNA. We established that treatment of the HL-60 cell culture with this peptide (10 ⁶ M) results in an increase in the number of apoptotic cells and suggested that HLDF is involved in processes of apoptosis.

Key words: apoptosis, differentiation factor HLDF, HL-60 cells, nuclease activity

Russian Journal of Bioorganic Chemistry, Vol. 26, No. 7, 2000, р . 450

A Biologically Active Fragment of the Differentiation Factor of the HL-60 Line Cells: Identification and Properties

I. A. Kostanyan*, ¹ M. V. Astapova*, E. V. Navolotskaya**, T. N. Lepikhova**, S. M. Dranitsyna*, G. B. Telegin**, I. L. Rodionov**, L. K. Baidakova**, Yu. A. Zolotarev***, I. M. Molotkovskaya*, and V. M. Lipkin*

*Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP-7 Moscow, 117871 Russia, **Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Pushchino Branch, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290 Russia, ***Institute of Molecular Genetics, Russian Academy of Sciences, Moscow

Abstract: Six-membered peptide fragment TGENHR (HLDF-6) was identified in the HL-60 cell culture of human promyelocyte leukemia treated with retinoic acid when studying the differentiation factor HLDF of this cell line. HLDF-6 retains the ability of the full-size factor to induce the differentiation and arrest the proliferation of the starting HL-60 cells. It was shown that the synthetic peptide HLDF-6 has no specific receptors on the surface of the HL-60 cells but can affect the binding of interleukin IL-1, a cytokine involved in proliferation, to the cell surface. It was found on a model of transplantable NSO myeloma that HLDF-6 has an antitumor activity.

Key words: cell differentiation factor, biologically active fragment; peptide; HL-60 cell line

Russian Journal of Bioorganic Chemistry, Vol. 26, No. 8, 2000, р . 505

The Biological Function of a Fragment of the Neurotrophic Factor from Pigment Epithelium: Structural and Functional Homology with the Differentiation Factor of the HL-60 Cell Line

I. A. Kostanyan*, ¹ S. S. Zhokhov*, M. V. Astapova*, S. M. Dranitsyna*, A. P. Bogachuk*, L. K. Baidakova**, I. L. Rodionov**, I. I. Baskin***, O. N. Golubeva****,J. Tombran-Tink*****, and V. M. Lipkin*

*Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP-7 Moscow, 117871 Russia, **Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Pushchino Branch, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290 Russia ***Chemical Faculty, Moscow State University, Vorob’evy Gory, Moscow, 119899 Russia, ****Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia *****Center for Neuroscience Research, Children’s National Medical Center and George Washington University, Washington, DC, USA

Abstract: It was shown that the full-size neurotrophic factor from pigment epithelium (PEDF) induces the cell differentiation of the human promyelocyte leukemia cell line HL-60. A structural analysis of PEDF revealed in its C-terminal region a six-membered peptide fragment PEDF-(352–357) (PEDF-6) whose sequence is highly homologous to the 41–46 fragment of the active site of the human leukocyte differentiation factor HLDF (HLDF-6). The biological effect of PEDF and synthetic peptides PEDF-6 and HLDF-6 on the HL-60 cells and the early gastrula ectoderm of Xenopus laevis embryos was studied. On the basis of the structural and functional homologies of HLDF, PEDF, and their homologous peptides and the computer models of the spatial structures of the full-size PEDF and the PEDF with the C-terminal fragment split off tby the cleavage of the Leu³⁸⁰–Thr³⁸¹ bond in the serpin loop, a hypothesis on the functional role of the serpin loop in PEDF was put forward.

Key words: cell differentiation; cell differentiation factors; peptides, computer modeling