LABORATORY OF PLANT MOLECULAR BIOLOGY
Head: Marek Figlerowicz
Research Staff:
Michał Jasiński,
Andrzej B. Legocki (President of Polish Academy of Sciences), Anna Modelska-Ziółkiewicz, Jan Podkowiński, Tomasz Stępkowski,
Paweł Stróżycki
Research Support:
Iwona Femiak, Lidia Jankowska, Hanna Glapiak
Ph.D. Students:
Magdalena Czaplińska, Błażena Grabowska, Agnieszka Kisiel, Katarzyna
Miedzińska, Anna Skąpska
Undergraduate Students:
Magdalena Bejger, Agnieszka Chlebicka, Izabela Ibragimow, Łukasz Markiewicz, Martyna Nowacka
Keywords:
plant genes, recombinant proteins, organ-specific gene expression, symbiotic
nitrogen fixation, plant transformation, ferritins, edible vaccines, RNA silencing, microRNA, DNA microarrays
Naturally occurring symbiotic systems between legumes and Rhizobium species
provide in-teresting models to study the controlling mechanisms of plant gene
expression. As a result of en-dosymbiotic interactions, nitrogen-fixing root
nodules are developed with significant alterations in gene expression of both
symbionts: host plant and microorganism.
The genetic material is prepared (cDNA and genomic libraries, cloned genes,
bacterial mutants) using the lupine plant-Bradyrhizobium sp. (Lupinus) symbiotic
system as a basic model. With that we are able to carry out detailed studies
on the expression of symbiotic genes at different stages of interaction using
molecular as well as cytological tools. Plant transformation systems provides
an important approach for testing the in vivo expression of recombinant DNA.
These systems were also adapted in our laboratory for engineering transgenic
plants producing medicinally important proteins.
Current research activities:
structure determination and expression analysis of plant symbiotic genes using
differential screening and array technology;
genetic analysis of nodulation region of B. sp. (Lupinus) controlling synthesis
of the Nod factor - a key morphogen for initiation of root nodule meristem;
phylogenetic and computation analyses of the plant genes and bacterial nodulation
genes to elucidate co-evolutionary correlations of symbiotic systems;
expression profiling of functionally linked genes for mRNA structure-function
relation with microarrays analysis;
iron metabolism, role of oxidative stress in legume symbiosis and molecular
links be-tween symbiotic and pathogenic interactions;
developing plant systems for expression of biopharmaceutically important proteins
in-cluding recombinant vaccines.
MOLECULAR VIROLOGY GROUP
Head: Marek Figlerowicz
Research Staff:
Józef Bujarski, Magdalena Alejska
Research Support: Nelli Malinowska
Ph.D. Student:
Paulina Jackowiak, Anna Urbanowicz
Undergraduate Students:
Renata Czołgosz, Anna Siek, Agnieszka Wojtkowiak
Keywords:
RNA viruses, retroviruses, genetic polymorphism of RNA viruses and
retroviruses, genetic RNA recombination,
error prone replication, viral replicase, reverse transctiptase, RNAi phenomenon,
viral vectors inducing RNAi,
DNA microarrays
Despite extraordinary progress that has recently been made in biological sciences
viral infections still remain one of the most serious problems in medicine
and agriculture. On the other hand, the unique features of viruses make them
a very interesting objects of basic studies and a useful tools in modern biotechnology.
The research conducted by the Team of Molecular Virology is especially focused
on RNA viruses and retroviruses. One of the most unusual futures of those
pathogens is their enormous genetic variability. It is the reason why RNA-based
viruses can easily adapt to environmental changes, escape from immunological
response or produce drug resistant mutants. At present, we have three systems
for investigating the genetic diversity in: brome mosaic virus (BMV), human
immunodeficiency virus (HIV) and hepatitis C virus (HCV).
During our studies we also try to take advantage from the fact that RNA viruses
infect, spread and express genetic information in higher organisms without
altering their genomes. As a result, there is a possibility that especially
prepared RNA virus-based vectors can be employed as useful tools in functional
genomics. Currently, we are testing two series of viral vectors: one is used
to silence selected host genes by inducing RNAi phenomenon and the other to
express certain genes in plant cells.
Current research activities:
- involvement of BMV replicase proteins in genetic RNA recombination;
- role of RNA structure in non-homologous recombination between genomic
molecules of BMV;
- homologous RNA recombination as a factor stabilizing the viral genome;
- determination of structural requirements for site-specific non-homologous
RNA recombination;
- cloning and expression of HIV reverse transcriptase its components and
mutants;
- reverse transcriptase as a key factor responsible for genetic variability
of HIV;
- cloning and expression of hepatitis B virus (HBV) reverse transcriptase
and hepatitis C virus (HCV) polymerase;
- determination of the mechanisms involved in the development of chronic
hepatitis C;
- BMV- and BBMV-based vectors inducing RNAi phenomenon in plant cells;
- RNAi phenomenon and DNA microarrays as effective tools in functional genomics;
- BMV- and BBMV-based vectors for the protein expression in plant cells.
SELECTED PUBLICATIONS
W. M. Karłowski, P. M. Stróżycki, A. B. Legocki
Characterization and expression analysis if the yellow lupin (Lupinus luteus L.) gene coding
for nodule specific prolin-rich protein.
Acta Biochim. Polon. 47, 371-383 (2000).
P. M. Stróżycki, W. M. Karłowski, Y. Dessaux, A. Petit, A. B. Legocki
Lupine leghemoglobin I - expression in transgenic Lotus and tobacco tissues.
Mol. Gen. Genet. 263, 173-182 (2000).
B. Łotocka, J. Kopcińska, W. Borucki, T. Stępkowski, A. Świderska, W. Golinowski, A. B. Legocki
The effects of mutations in nodZ and nolL genes of Bradyrhizobium sp. WM9 (Lupinus) upon the nodule structure in Lupinus luteus.
Acta Biol. Cracov. series Bot. 42, 155-163 (2000).
S. Wdowiak, W. Małek, E. Sajnaga, B. Łotocka, T. Stępkowski, A. B. Legocki
Symbiosis of Astragalus cicer with its microsymbionts: partial nodC gene sequence, host plant specificity, and root nodule structure.
Antonie Van Leeuwenhoek Journal of Microbiology 78, 63-71 (2000).
M. Figlerowicz
Role of RNA structure in non-homologous recombination between genomic RNA of brome
mosaic virus.
Nucleic Acid Res. 28, 1714-1723 (2000).
M. Figlerowicz, A. Bibiłło
RNA motifs mediating in vivo site-specific nonhomologous recombination in (+) RNA virus
enforce in vitro nonhomologous crossovers with HIV-1 reverse transcriptase.
RNA 6 339-351 (2000).
E. Bartoszak-Adamska, M. Figlerowicz, M. Wiewiórowski, T. Gustafsson, I. Olovson, M. Jaskólski
The influence of (O, N)-deuteration on the crystal and molecular structure, thermal stability and spectroscopic properties of 1,3-propanediammonium hydrogenphosphate monohydrate.
Polish J. Chem. 74, 393-408 (2000).
A. Bibiłło, M. Figlerowicz, K. Ziomek, R. Kierzek
The nonenzymatic hydrolysis of oligoribonucleotides VII. Structural elements affecting hydrolysis.
Nucleos. Nucleot. Nucl. 19, 977-994 (2000).
M. Alejska, A. Kurzyńska-Kokorniak, M. Broda, R. Kierzek, M. Figlerowicz
How RNA viruses exchange their genetic material.
Acta Biochim. Polon. 48, 391-407 (2001).
J. Kapusta, A. Modelska, T. Pniewski, M. Figlerowicz, K. Jankowski, O. Lisowa,
A. Płucienniczak, H. Koprowski, A. B. Legocki
Oral immunization of human with transgenic lettuce expressing hepatitis B surface antigen.
In: Advances in Experimental Medicine and Biology; Progress in Basic and Clinical
Immunology 495 Kluwer Academic/Plenum Publisher (2001).
T. Stępkowski, A. B. Legocki
Reduction of bacterial genome size and expansion resulting from obligate intracellular lifestyle and adaptation to soil habitat.
Acta Biochim. Polon. 48, 367-384 (2001).
E. Sajnaga, W. Małek, B. Łotocka, T. Stępkowski, A. B. Legocki
The root-nodule symbiosis between Sarothamnus scoparius L. and its microsymbionts.
Antonie van Leeuvenhoek Journal of Microbiology 79, 381-391 (2001).
K. Brzezinski, R. Janowski, J. Podkowinski, M. Jaskólski
Sequence determination and analysis of S-adenosyl-L-homocysteine hydrolase from yellow
lupine (Lupinus luteus).
Acta Biochim. Polon. 48, 477-83 (2001).
A. Wojtkowiak, A. Siek, M. Alejska, A. Jarmołowski, Z. Szweykowska-Kulińska,
M. Figlerowicz
RNAi and viral vectors as useful tools in the functional genomics of plants. Construction of BMV-based vectors for RNA delivery into plant cells.
Celullar and Molecular Biology Letters 7, 511-522 (2002).
A. Kurzyńska-Kokorniak, A. Urbanowicz, P. Jackowiak, A. Jankowska, N. Malinowska, M. Alejska, M. Figlerowicz
Construction of the universal system for the in vitro studies of HIV-1 reverse
transcriptase-mediated RNA recombination.
Annals of the Polish Chemical Society 2, 103-109 (2003).
M. Figlerowicz, M. Alejska, A. Kurzyńska-Kokorniak, M. Figlerowicz
Genetic variability – the key problem in prevention and therapy of RNA-based virus infections.
Medicinal Research Reviews 23, 488-518 (2003).
Z. Szweykowska-Kulińska, A. Jarmołowski, M. Figlerowicz
RNA interference and its role in the regulation of eucaryotic gene expression.
Acta Biochim. Polon. 50, 217-229 (2003).
T. Stępkowski, A. Świderska, K. Miedzinska, M. Czaplińska,
M. Świderski, J. Biesiadka, A. B. Legocki
Low sequence similarity and gene content of symbiotic clusters of Bradyrhizobium sp. WM9 (Lupinus) indicate early divergence of “lupin” lineage in the Bradyrhizobium genus.
Antonie Van Leeuwenhoek Journal of Microbiology 84, 115-124 (2003).
T. Stępkowski, M. Czaplińska, K. Miedzinska, L. Moulin
The variable part of the dnaK gene as an alternative marker for phylogenetic studies of rhizobia and related alpha Proteobacteria.
Systematic and Applied Microbiology 26, 483-494 (2003).
A. Kisiel, J. Podkowiński, A. B. Legocki
Normalised cDNA library from narrow-leaf lupine roots as a source of genes involved in symbio-tic interactions with Bradyrhizobium sp.
in Proceedings of the International Workshop “Understanding The Plant Genome” 69-78 (2003).
J. Podkowiński, J. Jeleńska, A. Sirikhachornkit, E. Zuther, R. Haselkorn, P. Górnicki
Expression of cytosolic and plastid acetyl-coenzyme A carboxylase genes in young wheat plants.
Plant Physiol. 131, 763-772 (2003).
K. Brzeziński, B. Rogoziński, T. Stępkowski, G. Bujacz, M. Jaskólski
Cloning, purification and preliminary crystallographic studies of Bradyrhizobium sp WM9 (Lu-pinus) fucosyltransferase NodZ.
Acta Crystallographica (series D) 60, 344-346 (2004).
L. Moulin, G. Béna, C. Boivin-Masson, T. Stępkowski
Phylogenetic analyses of symbiotic nodulation genes support vertical and lateral gene co-transfer within the Bradyrhizobium genus.
Molecular Phylogenetics and Evolution 30, 720-732 (2004).
D. Borek, K. Michalska, K. Brzeziński, A. Kisiel, J. Podkowiński, D. T. Bonthron, D. Krowarsch,
J. Otlewski, M. Jaskólski
Expression, purification, and catalytic activity of Lupinus luteus asparagine beta-aminohydrolase and its Escherichia coli homolog.
Eur. J. Biochem. 27, 3215-26 (2004).
E. Zuther, S. Huang, J. Jeleńska, H. Eilenberg, E. M. Arnold, X. Su, A. Sirikhachornkit,
J. Podkowinski, A. Zilberstein, R. Haselkorn, P. Górnicki
Complex nested promoters control tissue-specific expression of acetyl-CoA carboxylase genes in wheat.
Proc. Natl. Acad. Sci. USA 101, 1403-1408 (2004).
M. Barciszewska-Pacak, B. Grabowska, J. Wojciechowicz, A. Pacak, M. Figlerowicz
Virus-induced RNA silencing in plants.
In: Gene silencing by RNA interference, edited by M. Sohail, published by CRC Press, 323-343, (2004).
J. Błażewicz, M. Figlerowicz, P. Jackowiak, D. Janny, D. Jarczyński, M. Kasprzak, M. Nalewaj,
B. Nowierski, R. Styszyński, Ł. Szajkowiski, P. Widera
Parallel DNA Sequence Assembly.
In: Proceedings of the Fifth Mexican International Conference in Computer Science, edited by
R. Baeza-Yates, J. L. Marroquin and E. Chavez, published by IEEE Computer Society, 378-382 (2004).
J. Błażewicz, M. Figlerowicz, P. Formanowicz, M. Kasprzak, B. Nowierski, R. Styszyński,
Ł. Szajkowski, P. Widera, M. Wiktorczyk
Assembling the SARS-CoV genome – new method based on theoretic graph approach.
Acta Biochem. Polon. 4, 983-993 (2004).
T. Stępkowski, K. Brzeziński, A. B. Legocki, M. Jaskólski, G. Bena
Bayesian phylogenetic analysis reveals two-domain topology of S-adenosylhomocysteine
hydrolase protein sequences.
Molecular Phylogenetics and Evolution 34, 15-28 (2005).
A. Urbanowicz, M. Alejska, P. Formanowicz, J. Błażewicz, M. Figlerowicz, J. J. Bujarski
Frequent homologous crossovers among molecules of Brome mosaic bromovirus RNA1
or RNA2 segments in vivo.
Journal of Virology, in press.